FAQ

hi peiyong can i dilute concentrated lentivirus directly without filtering it first?
Yes, you can dilute concentrated lentivirus directly without filtering it first, provided that you maintain good lab practices to keep the virus stock sterile. Filtering is generally not necessary unless there are concerns about contamination during virus production or if you're working with high titer virus where large debris might be present. Just ensure to use an appropriate medium for dilution and handle everything in a sterile environment to minimize any risk of contamination.

hi peiyong how do i correctly dilute the concentrated lentivirus in pbs for my experiment?
To correctly dilute your concentrated lentivirus for your experiment, it's important to use a suitable medium rather than PBS. Since you mentioned that you've concentrated the virus by 1000 fold, you'll need to dilute it accordingly for testing. For instance, if you're using GoStix for detection, you should first dilute the concentrated virus 1000 times with the appropriate medium before proceeding with any further dilutions needed for your specific assay. This ensures accurate results in your experiments while maintaining the integrity of the viral sample.

what is the best strategy for optimizing virus yield in my packaging system using tet-free FBS?
To optimize virus yield in your packaging system using tetracycline-free FBS, consider the following strategies: 1. **Use Approved FBS**: Ensure that you are using Tet System approved FBS, specifically the TBUSA tetracycline-free serum, as it is functionally tested for compatibility with your lentiviral packaging vectors. 2. **Cell Passage Number**: Maintain your packaging cells within a passage range of 5-15 to ensure optimal growth and transfection efficiency. Avoid higher passages that may lead to reduced performance. 3. **Virus Collection Timing**: Collect the virus at approximately 48 hours post-transfection, as this timing aligns with typical practices in R&D for maximizing viral titer production. 4. **Monitor Genetic Insert Impact**: Acknowledge that the titer can vary based on the gene of interest (GOI). Therefore, be prepared to adjust parameters based on how different inserts affect viral yields. 5. **Avoid Contamination from Other Sources**: Be cautious about using serum from vendors other than those specified by Takara Bio, as they may contain unwanted tetracyclines which could interfere with expression systems. By implementing these strategies while adhering strictly to recommended protocols and materials, you should see improved outcomes in virus yield from your packaging system.

what degree dilution is needed to not fully inhibit pcr polymerases?
To avoid inhibiting PCR polymerases, it is recommended to dilute the reverse transcription (RT) reaction by a factor of 1/10. This dilution should help reduce the salt concentration and potentially improve compatibility with PCR enzymes like GXL or others.

is it possible to combine the enzymatic fragmentation module with the thruplex tag-seq flex kit while using UMIs in the thruplex adapters?
Yes, it is possible to combine the enzymatic fragmentation module with the ThruPLEX Tag-seq FLEX kit. The UMIs are incorporated using stem-loop adapters during the adapter ligation step, which are included in the kit. However, it's important to note that UMIs are not available for the ThruPLEX DNA-Seq kits; they are specifically designed for use with Tag-seq FLEX. If you need further assistance or have more questions about your setup, feel free to ask!

is it possible to combine the enzymatic fragmentation module with thruplex tag-seq and use UMIs present in thooplex adapters?
It is not possible to combine the enzymatic fragmentation module with ThruPLEX Tag-seq and use UMIs from ThruPLEX adapters, as UMIs are only available for the Tag-seq FLEX. The ThruPLEX DNA-Seq products do not include UMIs. If you have further questions or need assistance with a specific application, feel free to ask!

how can I achieve higher functional titer for lentix cells using the current protocol?
To achieve a higher functional titer for Lenti-X cells using your current protocol, consider the following strategies: 1. **Plasmid Quality**: Ensure that your plasmid DNA is of high purity and integrity. Aim for 260/280 and 260/230 ratios around 2 or better. 2. **Transfection Efficiency**: Strive for transfection efficiencies greater than 90%. This can be optimized by adjusting the amount of transfecting agent used and ensuring optimal cell health prior to transfection. 3. **Cell Condition**: Use low passage 293T cells grown in Tet-approved FBS, as these conditions have been shown to support improved viral production. Make sure the cells are at about 60-70% confluence before starting transfection. 4. **Serum Usage**: Utilize Tet-approved FBS two passages prior to transfection if you haven't already done so, as this may impact cell viability and productivity. 5. **Vector Size Considerations**: Confirm that the total size from LTR to LTR does not exceed 9.7 kb; larger constructs may lead to unstable viral particles and reduced titers. 6. **Collecting Supernatant Timing**: Optimize when you collect the supernatant post-transfection; typically collecting it after around 48-72 hours can yield better results, but this might vary based on specific protocols or vector systems employed. 7. **Increasing Transgene Copy Number**: If applicable, increasing the copy number of your lentiviral plasmids during packaging could enhance overall titer output. By focusing on these areas within your existing protocol framework, you should see improvements in functional titers from your lentiviral preparations.

what would be the best library prep kit for analyzing total rna from extracellular vesicles with low input, considering the differences between smart-seq and nebnext kits?
For analyzing total RNA from extracellular vesicles (EVs) with low input, the SMART-Seq® Total RNA Pico Input with UMIs (ZapR® Mammalian) is a strong option. This kit is designed to handle small amounts of starting material and incorporates unique molecular identifiers (UMIs), which can enhance data accuracy by distinguishing between PCR duplicates. In contrast, while NEBNext kits are also effective for library preparation, they may not be specifically optimized for single-cell applications or extremely low-input scenarios like those involving EV RNA. Additionally, it’s crucial to ensure that any single-stranded DNA contamination is removed prior to cDNA synthesis, as this could significantly impact results. It's advisable to consider your specific research needs and experiment parameters when making a final decision on the library prep kit. The cited literature may provide further insights into performance differences between these options in similar contexts.

what's the best way to effectively remove genomic dna from my rna samples using this kit?
To effectively remove genomic DNA from your RNA samples using the NucleoSpin RNA kit, it is recommended to include a DNase treatment step during the purification process. This involves adding DNase enzyme to your RNA sample after lysis and before the binding step. Follow the kit's instructions for optimal conditions regarding incubation time and temperature for the DNase treatment. After completing the initial RNA isolation, consider performing an additional clean-up using the NucleoSpin® RNA Clean-up XS kit. This will help ensure that any residual DNase is removed, along with impurities that may affect downstream applications. By combining these steps—using effective DNase treatment followed by cleanup—you can significantly enhance the purity of your RNA samples while minimizing any potential contamination from genomic DNA.

know nucleomag dna rna pro work different kit you'd recommend?
The NucleoMag® DNA/RNA kit is designed for the isolation of both DNA and RNA from various samples, including blood. If you're looking for alternatives, the NucleoSpin RNA + DNA/RNA Buffer set could be a good choice if your focus is on RNA extraction specifically. For blood samples, you might consider using the NucleoSpin Blood L or XL kits as they are optimized for extracting nucleic acids from whole blood. Always consult the manufacturer’s recommendations to ensure compatibility with your sample type and desired output.

Is there a manual available for the Mupid Plus Include Gel Maker Set Large?
It would be best to check the manufacturer's website or contact their customer support for a manual specific to the Mupid Plus Include Gel Maker Set Large. They often provide downloadable manuals or can assist you with any questions you might have regarding the product. If you're unable to find it online, I can help look for additional resources after a search.

what are the dimensions of the plate?
The dimensions of the plate are currently unavailable in the database. It would be best to check back later, as additional sources may provide the information needed. An email will be sent once more details have been found. Thank you for your understanding!

do i need to include a dnase digestion step during the rna isolation process with takara's nucleospin rna virus kit?
Including a DNase digestion step during the RNA isolation process with Takara's NucleoSpin RNA Virus kit is recommended, especially if your application requires DNA-free RNA. While the kit is designed to minimize gDNA contamination, adding a DNase treatment can further ensure that any residual DNA is removed. You can perform this step after isolating the RNA by following the appropriate protocol and using an RNase-free DNase enzyme. Just make sure to clean up the RNA afterward to remove any remaining DNase that could potentially interfere with downstream applications like ddPCR or RT-qPCR.

kit help need know amplify rnas samples synthetize cdna?
To amplify RNA samples and synthesize cDNA, you can use the CellAmp™ Whole Transcriptome Amplification Kit (Real Time), Ver. 2. This kit is specifically designed for amplifying cDNA from trace amounts of total RNA, allowing effective pre-amplification before proceeding with qPCR. You can find detailed instructions in the user manual available on the Takara Bio website, particularly on page 8 where the protocol is outlined. If you're looking to convert your RNA into cDNA directly without pre-amplification, consider using the PrimeScript™ RT Reagent Kit (Perfect Real Time), which requires a minimum of 10-20 ng of total RNA for optimal results but does not support amplification prior to conversion.

do you sell silica-based dna extraction kits for kingfisher instruments?
The NucleoMag kits mentioned are compatible with KingFisher platforms and provide options for genomic DNA purification from tissue samples. While the specific formulation details of the beads used in these kits, including whether they are silica-based, are not disclosed by MN, they are comparable to Ampure beads. If you're looking specifically for silica-based DNA extraction kits, it's best to check directly with suppliers or manufacturers that specialize in such products for compatibility with KingFisher instruments.

how prepare rna extraction protocol for limited cell number sorted tumor cells from lung mice using direct capture method
To prepare an RNA extraction protocol for limited cell number sorted tumor cells from lung mice using the direct capture method, follow these steps: 1. **Cell Sorting**: Begin by sorting your tumor cells into PBS to minimize contamination with the sorting buffer. Ensure that you are aware of the number of cells being sorted. 2. **Pelleting Cells**: After sorting, centrifuge the cells at a low speed (e.g., 300-500 x g for 5-10 minutes) to form a pellet. 3. **Lysis Buffer Preparation**: Carefully aspirate the supernatant without disturbing the pellet and resuspend it in Lysis Buffer LB1 provided in your NucleoSpin® RNA Plus XS kit. 4. **RNA Extraction Procedure**: - Follow the manufacturer’s instructions outlined in the NucleoSpin® RNA Plus XS manual for cell lysis and subsequent purification steps. - Make sure to include any additional reagents or protocols specific to handling small numbers of cells. 5. **Quality Check**: Once extracted, assess RNA quality and quantity using spectrophotometry or other suitable methods before proceeding with downstream applications such as qPCR or sequencing. By following this structured approach, you'll ensure optimal recovery and integrity of RNA from your limited cell numbers post-sorting.

does the smart-seq mrna lp kit typically come with aliquoted nucleomag ngs clean-up beads and can ampure xp beads be used as a suitable alternative?
The SMART-Seq mRNA LP kit does not typically come with aliquoted NucleoMag NGS Clean-up and Size Select Beads. While AMPure XP beads can be used as a suitable alternative, it is important to follow the recommended bead:sample ratios for the NucleoMag process. Additionally, only use AMPure XP beads from the specified catalog numbers (A63880 & A63881) if necessary, as they have been validated for this workflow. Avoid using any other types of SPRI beads.

what is the status of my order?
To check the status of your order, it's best to contact customer service directly. You can reach them by calling their dedicated line or emailing them for assistance. They will be able to provide you with the most accurate and up-to-date information regarding your order status. If you need help with anything else, feel free to ask!

what kit sound need?
For your application in qPCR or PCR, the PrimeScript™ RT Reagent Kit (Perfect Real Time) would be a suitable choice. This kit is highly sensitive and has been proven to work effectively with as little as 10 pg of total RNA, making it ideal for your needs if you're working with 10-20 ng of total RNA. If you have your own oligos, you can also opt to purchase just the reverse transcriptase separately. For more detailed information, you can refer to the user manual provided by Takara Bio.

why is there no gfp signal in my transfected lentiviral cells?
The absence of GFP signal in your transfected lentiviral cells could be due to several factors, particularly related to the use of an IRES element. If there is no gene upstream of the IRES, it can lead to poor expression of the downstream gene, which in this case is likely the GFP. This might delay fluorescence visibility beyond your normal observation period after transfection or transduction. Typically, with regular GFP vectors, you would expect to see fluorescence within about 48 hours post-transfection; however, when using an IRES without an upstream insert—or even with one that isn't effectively expressed—the time frame for observing any signal can extend significantly. Testing your vector with the insert should help confirm whether this hypothesis holds true and provide insights into optimizing expression levels moving forward.

what options do you have that aren't random?
For non-random mutagenesis and cloning, you might consider using directed mutagenesis techniques or methods like Gibson Assembly. The In-Fusion method is also a great option as it allows for the integration of inserts at specific locations within a plasmid. These techniques can provide precise control over DNA modifications compared to random approaches often associated with transposon-based systems. If you have specific requirements in terms of insert size or target location, that could further guide your choice among these options.

please provide expiration date for thiophilic superflow resin
The expiration date for thiophilic superflow resin can vary based on the manufacturer and specific product batch. It's best to check the product packaging or contact the supplier directly for accurate information regarding its shelf life. If you need further assistance, feel free to ask!

how can i get help with the gif?
To get help with the gif, you might consider checking if there are any specific guidelines or resources available in your workspace related to using gifs. Additionally, reaching out to a colleague who is familiar with the platform could provide clarity on how to effectively use and share gifs. If you're looking for technical support, contacting your IT department or support team may also be beneficial.

can we try again with the alerter?
It sounds like the alerter has worked for you and Bakhyt, which is a good sign. Since Marianne was away from her table, it might be helpful to check in with her when she’s back to confirm whether it’s working on her end as well. If everyone is ready, we can definitely try again with the alerter to ensure everything functions smoothly.

how can i troubleshoot my primers for better binding and amplification of the plasmid?
To troubleshoot your primers for better binding and amplification of the plasmid, consider extending the 3' end of the primer to increase specificity, especially if you are working with repeat sequences. This can help achieve a more targeted amplification. Additionally, utilize the Primer Design Tool available on our website by entering your sequence information. If you encounter issues, please click on "Report Issues/Ask Question" at the bottom of the page to submit feedback about your primer design file. It is also helpful to send us details about the template vector you are using for amplifying the insert so that we can provide more tailored assistance.

what recommended protocols products specifically for viral transduction in murine cancer cell lines?
For viral transduction in murine cancer cell lines, using a retroviral packaging system is effective. You can consider employing fluorescent protein-expressing retroviral vectors from reliable sources like Takara Bio, which offer various options for gene delivery. It’s essential to ensure that the targeted cell line is amenable to transduction. If you're working with cells that are easier to transfect rather than transduce, you might want to use a reagent such as Xfect for your experiments. The Xfect protocol provides detailed instructions on how to perform the transfection efficiently. For optimal results in both approaches—transfection and transduction—it’s important to know specific characteristics of your target cells, including their proliferative status and any unique requirements they may have. If there are particular challenges or needs regarding your experimental setup or cell type, feel free to explore tailored solutions based on those factors.

could you provide details on the availability of smart-seq kits for mouse TCR and UMIs?
The SMART-Seq® Mouse TCR (with UMIs) is currently available for purchase. For this kit, it's important to note that the Unique Dual Index (UDI) Kit must be purchased separately. The UDI kits allow for preparing and pooling up to 384 Illumina-compatible libraries, with options available based on the number of libraries you plan to pool. For further details or a quote, it would be best to connect with Sales directly. They can provide specific pricing and availability information tailored to your needs. If you have any additional questions or require technical support, please let me know!

can you provide the exact size of groEL protein?
The groEL protein is composed of 549 amino acids, which corresponds to a nucleotide sequence length of 1647 nucleotides, as it is located between nucleotides 2154 and 3800 in the provided vector sequence.

What is the status of my order?
The status of your order can be checked by reaching out to our account handling team, who will provide you with the most accurate and up-to-date information. If there's anything else you'd like assistance with, feel free to ask!

is there a more affordable option that matches the performance of this kit?
There may be more affordable options available that can match the performance of the TB Green® Advantage® qPCR Premix. It's worth exploring other brands or formulations that offer similar features, such as efficiency and sensitivity in real-time PCR applications. Additionally, checking for bulk purchase discounts or promotional offers could help reduce costs. Consider reaching out to suppliers for recommendations on budget-friendly alternatives that meet your specific needs while maintaining quality performance.

what if want confirm effective transduction test using different values?
To confirm effective transduction using different values, you can perform a series of experiments varying the multiplicity of infection (MOI) based on your lentiviral preparations. Start by calculating the IFU using the GoStix or any other titer kit to determine your current viral concentration in terms of ng/ml of p24. Once you have that information, adjust your MOI by testing various dilutions of your virus preparation. For example, if you're starting with a GV value indicating 500 ng/ml of p24, which corresponds roughly to an upper range around 10e7 IFU/ml, consider preparing serial dilutions to achieve lower and higher MOIs. Transduce target cells with these different dilutions and monitor transduction efficiency through a suitable readout method—such as flow cytometry for GFP expression or another reporter gene specific to your vector system. This approach allows you to identify the optimal MOI for effective transduction while ensuring that each experimental condition is well-controlled. After performing these tests, analyze how changes in input viral load affect outcomes like transgene expression or cell viability. Over time and across several preps, you'll gain insights into consistent patterns that will guide future experiments more confidently without needing extensive recalibrations every time.

how should i optimize the process for storing and managing my cell library effectively?
To optimize the process for storing and managing your cell library effectively, consider the following steps: 1. **Use Glass Beads**: Start by placing 15 sterilized glass beads in the center of each plate to facilitate even distribution when freezing. 2. **Gradual Freezing**: Stack 5-10 plates on top of each other and place them on a shaker to ensure gradual freezing rather than flash-freezing, which can damage cells. 3. **Aliquoting**: After achieving proper freezing, aliquot your samples and transfer them to -80°C for long-term storage. 4. **Careful Handling**: When working with cells, avoid using forceful spreaders that may smash the cells; instead, opt for gentler methods or manual techniques while ensuring not to spill any contents. 5. **Monitoring Colony Counts**: Regularly check the number of individual colonies formed during cultivation to assess viability and growth rates. 6. **Check Titer Levels**: Utilize a densitometer periodically to measure titer levels; aim for values around <2 x 10^7 per ml as an indicator of optimal concentration. By incorporating these practices into your workflow, you will enhance both the preservation and management of your cell library efficiently.

anything improve titer?
To improve titer, consider the following suggestions: 1. **Cell Health and Density**: Ensure that your packaging cells are healthy and maintained between 20% to 80% confluency. Increasing the plating density to around 80-90% at the time of transfection may enhance your results. 2. **Use of Endotoxin-Free DNA**: If you haven't already, switch to endotoxin-free plasmid DNA preparation for transfections. Endotoxins can be detrimental to mammalian cell health, impacting viral production negatively. 3. **Passage Number**: Stick with packaging cells in passages 5 through 25 after thawing, as they generally perform better. 4. **FBS Quality**: Use tet-free FBS during packaging if it's not already being used, as this can positively influence cell growth and viability. 5. **Plasmid Backbone**: Since you're using a standard third-generation SIN lenti plasmid backbone which has been confirmed fine, ensure all components are correctly optimized for your specific application. 6. **Monitor Doubling Time**: Keep an eye on the doubling time of your cells; slower-growing populations might affect overall yield negatively. By implementing these strategies, there is a good chance you'll see improvements in your titers without too much additional work involved in redoing procedures like plasmid prep immediately.

need help finding expiration date for these products?
To find the expiration dates for the products, it's best to check the lot numbers on their packaging. If you have the catalog number of the kit, that can also be helpful in searching for specific information. It seems there are concerns about some older buffers, including 10X Titanium Taq PCR buffer and potentially others like Ex Taq. Since these products may be quite old and possibly expired, contacting the manufacturer directly would be a reliable way to obtain accurate expiration data. If you're unable to locate a lot number or need further assistance with this process, reaching out to customer service or checking product documentation can also provide valuable insights.

recommend protocol plant samples using mightyprep and terra with primestar gxl?
To extract DNA from plant samples using MightyPrep and then perform PCR with PrimeSTAR GXL, follow these steps: 1. **Sample Preparation:** - Choose your plant tissue (leaves are preferable as they lyse more easily than roots or other tissues). - Cut a small piece of the leaf sample to use for extraction. 2. **DNA Extraction Using MightyPrep:** - Add 100 μl of MightyPrep reagent for DNA into a 1.5-ml tube. - Use sterile tools to transfer the small leaf sample into this tube, ensuring it is well-resuspended in the reagent. - Follow Steps 3-5 outlined in the MightyPrep user manual to complete the lysis and subsequent purification process. 3. **PCR Amplification with PrimeSTAR GXL:** - Once you have your purified DNA extract, prepare your PCR reaction according to PrimeSTAR GXL's protocol. - Typically, this involves combining template DNA, primers specific to your target sequence, dNTPs, buffer solution, and PrimeSTAR GXL polymerase in appropriate volumes. 4. **Thermal Cycling Conditions:** - Set up thermal cycling conditions based on your target amplicon size and primer specifications; ensure that annealing temperatures are optimized for effective amplification. By following this protocol using MightyPrep for extraction followed by PCR with PrimeSTAR GXL, you should achieve reliable results suitable for genotyping purposes without requiring high fidelity due to no critical need mentioned in your context.

what's the best approach to optimize yield while considering cost and copy number?
To optimize yield while considering cost and copy number, it's important to carefully balance the amount of cell pellet used with the specific requirements of your experiment. Begin by determining the optimal cell density for your application, as this can significantly impact both yield and cost-effectiveness. Consider conducting a series of small-scale tests to identify the ideal conditions that maximize output based on varying copy numbers. This will allow you to assess how different parameters affect yield without incurring high costs. Additionally, ensure you're using an appropriate concentration of reagents from your MIDI kit tailored to your setup, as using too much or too little can lead to inefficient results. Keeping thorough records of each trial will help in fine-tuning your approach over time. Lastly, review any available literature or protocols related to similar experiments that may provide insights into optimizing yields effectively within budget constraints.

could using a heat block instead of a thermocycler improve my colony formation and help with the annealing process?
Using a heat block instead of a thermocycler could potentially improve your colony formation and assist with the annealing process. A slow ramping time in a thermocycler may prolong the digestion of DNA ends, which can hinder proper annealing and repair during cloning. Preheating the heat block to 50°C for incubation might minimize this issue by allowing you to control the temperature more effectively without extended exposure that could degrade your overlapping sequences. It's worth trying out this method to see if it enhances your results, as feedback from other users indicates it may be beneficial.

what do you think causes the color difference between the two ypd batches?
The color difference between the two YPD batches could be attributed to several factors, including variations in the raw materials used, differences in the preparation process, or even slight changes in environmental conditions during production. Since it's a rich medium, small scale transformations can also lead to noticeable shifts in color. Ensuring consistency in ingredient sourcing and processing methods may help minimize these variations.

what options do i need to select for in-fusion fusion cloning primer design without preserving restriction sites?
When designing primers for In-Fusion cloning without the need to preserve restriction sites, you can follow these guidelines: 1. **Select Overlap Regions**: Choose primer sequences that include overlapping regions with your vector and insert. The overlap should typically be around 15-20 base pairs. 2. **Avoid Restriction Sites**: Ensure that your chosen primer sequences do not contain any restriction enzyme recognition sites, as these will not be preserved in the final construct. 3. **Include Fusion Tags (if needed)**: If you are generating a fusion protein, consider adding coding sequences for any necessary tags at the ends of your primers to facilitate downstream applications like purification or detection. 4. **Consider Melting Temperature (Tm)**: Design your primers so they have similar melting temperatures to ensure efficient annealing during PCR amplification. 5. **Check Specificity**: Use software tools or online databases to confirm that your designed primers are specific to the target sequence and won't bind non-specifically. By following these steps, you can effectively design In-Fusion cloning primers tailored to your needs without preserving restriction sites while ensuring successful cloning outcomes.

what is the maximum dna size I can obtain using downstream nucleomag nucleobond hmw kits?
The NucleoBond® HMW kits are designed for the purification of high molecular weight DNA, typically allowing you to obtain fragments up to several hundred kilobases in size. For specific maximum sizes or more detailed information regarding your particular application, it's best to refer to the product documentation or consult with technical support directly.

what is the maximum dna size I can expect from the nucleomag nucleobond hmw kits?
The NucleoBond HMW kits are designed for the purification of high molecular weight DNA, and you can expect to obtain DNA fragments that are typically in the range of several hundred kilobases up to 1 megabase or more. However, specific results may vary depending on your sample type and conditions used during purification. For optimal outcomes, it's recommended to follow the kit's guidelines closely. If you're working with genomic DNA cleanup instead, consider exploring other specialized kits available for that purpose.

what's consequence of lower efficiency adapter ligation mean rare transcripts not amplified?
Lower efficiency in adapter ligation can lead to a diminished representation of rare transcripts in the final sequencing library. This is because if the adapters are not efficiently attached to all RNA molecules, those that do not have adapters will not be amplified during subsequent PCR steps. As a result, any rare transcripts present may be underrepresented or completely missed in the sequencing data, potentially leading to incomplete insights into gene expression profiles and missing critical biological information. It's essential to ensure efficient library preparation methods to capture the full range of transcript abundance accurately.

why is there such a big price difference between ampure xp and nucleomag size select beads?
The price difference between Ampure XP and NucleoMag Size Select beads can be attributed to several factors. Firstly, the manufacturing processes and quality control standards may differ between the two products, impacting their production costs. Additionally, the specific applications for which each bead type is optimized could influence pricing; if one product is designed for a broader range of uses or offers unique features, it might command a higher price. Furthermore, brand reputation and market demand play significant roles in pricing strategies. If one product has established itself as more reliable or efficient within certain workflows—like the SMARTer® smRNA-Seq Kit—the cost may reflect that perceived value among users. Ultimately, it's important to consider these aspects when evaluating options for your experiments to ensure you choose the right fit based on both performance needs and budget constraints.

What is the expiration date for the part number in the ERP system?
The expiration date for the part number in the ERP system is January 31, 2026, based on lot number AO8P027.

does the co-ip kit contain detergent?
Based on your description, it seems likely that the co-immunoprecipitation (co-IP) kit may contain a small amount of non-ionic detergent, especially if it's designed to maintain protein interactions while minimizing disruption. It's always best to check the product specifications or datasheet for detailed information about the components included in the kit.

hello marianne could you confirm if the new ss-total pico kit is compatible with our existing workflow and what adjustments we might need to make?
The new Smart-Seq Total RNA Pico Input kit is designed to be compatible with existing workflows, particularly those utilizing unique dual index kits. However, adjustments may be necessary depending on your specific setup and protocols. It would be beneficial to review the user manual for detailed compatibility information and instructions related to sample input amounts, library preparation steps, and any modifications required for optimal performance in your workflow. If you have specific components or methods currently in use, comparing them against the guidelines provided by the manufacturer will help ensure a smooth integration of the new kit into your processes.

what suggestions improve titer?
To improve titer, consider the following suggestions: 1. **Cell Health and Plating Density**: Ensure that the cells are healthy and properly plated. Culturing cells immediately after receiving them may not yield optimal results if they aren't in good condition. Allowing the cells to go through a few passages can help ensure they are robust before transfection. 2. **Coating Plates with Collagen**: Coating plates with collagen can enhance cell adherence, which often leads to improved health and transfection efficiency by providing better surface area for cell attachment. 3. **Adjusting Plating Densities**: Experiment with different plating densities to identify the optimum conditions for your specific transfection process. 4. **Using LentiX Concentrator**: If low titers persist, using a LentiX concentrator can help increase viral titer effectively. 5. **Cargo Size Consideration**: Be mindful of the size of the gene being cloned into your vector; larger genes (around 3kb or more) may negatively impact both titer and mCherry expression due to limited cargo space in the vector. By implementing these strategies, you should see an improvement in viral titer during your experiments.

know can you explain why sometimes transduction efficiency varies and what factors might influence the success of retroviral supernatant in different temperatures?
Transduction efficiency can vary due to several factors, particularly when using retroviral supernatants. One primary factor is the temperature during spinoculation or incubation. When cells are exposed to lower temperatures (like 4°C), their membranes may become less permissive for viral entry compared to warmer conditions (25-32°C). This reduced permeability can lead to decreased transduction efficiency. Other influencing factors include: 1. **Virus Titer**: The concentration of infectious units in your viral preparation is crucial. It's important to compare your titer with a control and ensure you're measuring equivalent units, such as infectious units per milliliter (IFU/ml). 2. **Cell Type**: Different cell lines have varying susceptibilities to infection based on their receptor expression and membrane characteristics. 3. **Binding Time**: A shorter binding time during centrifugation might not allow sufficient interaction between the virus and the cells, which could limit effective transduction. 4. **Viral Preparation Method**: Concentrated preparations versus standard ones may yield different efficiencies due to variations in how well they maintain infectivity. 5. **Use of Coating Agents like RetroNectin**: The method used for coating wells influences how effectively viruses bind and enter target cells, especially if protocols deviate from recommended guidelines. To optimize transduction efficiency, consider performing pilot experiments under various conditions—such as testing different temperatures or methods—and refer back to user manuals for specific protocols tailored for your applications.

sure page receiving kits replacement week please send email confirmation provide pi?
A replacement kit will be processed once the necessary details are confirmed with customer support. You can expect an email confirmation regarding the shipping status and any relevant information related to your order. If you need a purchase order number, it will be created after this chat session concludes. Thank you for your patience!

what are typical concentrations of Tris, salt, and metal ions for restriction digest in a reaction involving glycerol?
In a restriction digest reaction involving glycerol, typical concentrations for the components can vary. For Tris, concentrations are usually around 10-15 mM. Salt concentration may be approximately 50 mM to help maintain ionic strength and proper enzyme activity. Metal ions such as Mg²⁺ are commonly included at concentrations ranging from 1 to 10 mM, depending on the specific requirements of the restriction enzymes used. However, it’s important to note that the presence of glycerol can interfere with reactions, so optimizing conditions based on your specific system is advisable.

how can i access my order without account access?
If you need to access your order without account access, the best approach is to contact customer support directly. You can reach them by calling 800.662.2566 and selecting option 1 for assistance. They are available Monday through Friday from 6:00 am to 5:00 pm Pacific time and will be able to help you with your order inquiries or any other concerns you may have.

am us am looking for bioinformatics position openings in companies in Japan, can you help me find job links?
While I can’t provide specific job links for bioinformatics positions in Japan, I recommend checking company websites directly or using job search platforms that cater to the Japanese market. You may also consider networking on professional sites like LinkedIn or reaching out to recruitment agencies that specialize in life sciences and technology roles in Japan. If you're interested in opportunities at Takara, you can visit their careers page for openings specifically available in the US.

any other tips or best practices for optimizing lentivirus production that you've seen work well?
To optimize lentivirus production, consider the following best practices: 1. **Cell Health and Density**: Ensure that your producer cells are healthy and at optimal confluency. Passaging them to ensure robust growth can significantly impact virus yield. 2. **Coating Plates**: Using collagen or other extracellular matrix components to coat culture plates can enhance cell adhesion and flattening, which may improve transfection efficiency. 3. **Transfection Reagents**: Experiment with different transfection reagents as their efficacy can vary depending on the specific cell line used. 4. **Optimizing DNA Ratios**: Adjust the ratio of transfer plasmid to packaging plasmids during transfection; sometimes a higher proportion of transfer plasmid can lead to better yields. 5. **Incubation Conditions**: Maintain optimal incubation conditions such as temperature, CO2 levels, and humidity during both transfection and post-transfection phases for improved viral production. 6. **Use of Concentrators**: If titers remain low, employing a lentiX concentrator or similar methods post-production can help concentrate the virus without losing significant functionality. 7. **Adjusting Plating Densities**: Testing various plating densities might reveal an optimal density for maximal productivity based on your specific cell type and experimental setup. 8. **Monitoring Time Points**: Harvest supernatants at multiple time points after transduction to identify when peak viral titers occur for future experiments. By implementing these strategies alongside careful monitoring of each step in the process, you should see improvements in lentivirus production efficiency.

what is the concentration of rock inhibitor in takara cellartis beta cell kit supplement?
The concentration of the ROCK inhibitor in the Takara Cellartis beta cell kit supplement is not disclosed. While R&D may provide information on whether the ROCK inhibitor is included, they typically do not share specific concentration details. If you need further assistance or updates, please await a response via email from our team.

what are the key differences between tet system approved fbs us-sourced and non-us sourced tet system approved fbs?
The key differences between US-sourced and non-US sourced Tet system approved FBS primarily lie in their source, as there may be variations in regulatory standards, quality control processes, and potential contamination risks associated with different geographical origins. However, in terms of performance for cell culture applications using the Tet system, there are typically no significant differences noted. Both types can provide similar results when used appropriately within the context of your experiments. It's always best to evaluate based on specific needs and any available data on performance outcomes for your particular application.

what should i do with the thawed dry ice package now?
If you have a thawed dry ice package, it's important to handle it safely. First, ensure that the area is well-ventilated, as dry ice sublimates into carbon dioxide gas. If the dry ice is still in solid form but melting, you can leave it at room temperature until it completely evaporates. Do not touch the dry ice directly with your bare hands; use gloves or tongs to avoid skin burns. If there are any specific disposal instructions from the supplier or manufacturer, be sure to follow those guidelines as well.

where can i find additional documentation for retro-x retro vector set?
Additional documentation for the Retro-X Q Vector Set can be found through the following links: 1. The Certificate of Analysis can be accessed here: [Certificate of Analysis](https://www.takarabio.com/documents/Certificate%20of%20Analysis/631516/631516.pdf). 2. For a comprehensive overview, you can refer to the Retroviral Gene Transfer and Expression User Manual available at this link: [User Manual](https://www.takarabio.com/documents/User%20Manual/Retroviral%20Gene%20Transfer%20and%20Expression%20User%20Manual/Retroviral%20Gene%20Transfer%20and%20Expression%20User%20Manual.pdf). These resources should provide you with detailed information regarding the vector set.

what should i do if my pellet is large and white?
If your pellet is large and white, it may indicate a high concentration of proteins or other materials. It's important to proceed with caution to avoid disturbing the pellet during centrifugation. Make sure you're using appropriate speed and time settings for your specific protocol. If needed, you can mark the tube where the pellet will collect to help guide your pipetting when removing supernatant. Additionally, if any precipitated protein in the media helps you locate the pellet more easily, that could be beneficial as well. If you're still encountering issues or have concerns about the quality of your sample, consider reviewing your protocol or seeking additional guidance on handling larger pellets effectively.

how do i prepare the cds fragment of my gene of interest for mRNA synthesis?
To prepare the CDS (coding sequence) fragment of your gene of interest for mRNA synthesis, you'll need to start by PCR amplifying your target sequence using appropriate primers. It's important to design these primers with specific features that facilitate cloning, as outlined in the user manual you referenced. Once you've amplified the CDS fragment, you will clone it into a template vector using In-Fusion cloning. This process allows for seamless integration of your PCR product into the vector. Make sure to follow the guidelines provided in Section VI of the user manual closely. After successfully cloning your CDS into the vector, you can then proceed with mRNA synthesis from this construct following standard protocols for transcription. If you have further questions or need additional assistance with any step in this process, feel free to ask!

What are the features of your lentivirus purification kits?
Lentivirus purification kits typically offer several key features designed to facilitate the efficient and effective isolation of lentiviral particles. These may include: 1. **High Purity**: The kits are engineered to provide high-purity viral preparations, minimizing contaminants that could interfere with downstream applications. 2. **Ease of Use**: They often come with streamlined protocols that simplify the purification process, making it accessible even for users with limited experience. 3. **Scalability**: Many kits allow for flexibility in terms of scale, accommodating small laboratory batches as well as larger production needs. 4. **Fast Processing Time**: The purification procedures are generally optimized to reduce processing time without compromising yield or quality. 5. **Compatibility**: Kits are usually compatible with various cell types and can be used for different applications, including gene delivery and functional studies. 6. **Reproducibility**: They are designed to produce consistent results across multiple experiments, which is crucial for research reliability. These features collectively help researchers efficiently purify lentiviruses while ensuring high-quality outcomes in their experimental work.

what is the shelf-life of lenti-x lenti concentrator?
The shelf life of Lenti-X Lenti Concentrator is typically around 12 months when stored properly. However, if it has been opened or is close to 7 years old, as mentioned in the conversation, it's advisable to order a new bottle for optimal results. Proper storage and keeping the lid tightly sealed can help prevent issues like evaporation.

when can i get tet-fbs to culture cells immediately?
Tet-free FBS is typically used when introducing a vector with a TRE plasmid for gene expression. Until that point, regular FBS should be sufficient for culturing cells. If you are planning to transition to using tet-free FBS specifically for your experiments, it would be best to obtain it shortly before you need to introduce the vector into your cell cultures. This way, you can ensure optimal conditions right from the start of your gene expression work.

what are the differences between isolating cells using your entire cell population kits and other methods?
Isolating cells using entire cell population kits typically offers several advantages compared to other methods. These kits are designed to efficiently capture a broad range of cell types, ensuring that you obtain a representative sample from the starting material. This can be particularly beneficial when studying heterogeneous populations. In contrast, other methods may focus on isolating specific cell types or subpopulations, which could lead to bias if those targeted cells are underrepresented in your sample. Additionally, entire cell population kits often provide streamlined protocols for quick and reproducible results, minimizing hands-on time and variability between experiments. Moreover, these kits might include features such as optimized buffers and reagents that enhance cell viability and yield during isolation processes. Overall, using an entire cell population kit allows for a more comprehensive analysis of cellular behavior while maintaining integrity across various applications.

what suitable lentiviral vector can be used to express flavivirus envelopes for pseudovirus neutralization assays?
For expressing flavivirus envelopes in pseudovirus neutralization assays, you can use the lentiviral vectors available from your product line. Specifically, the single shots without envelope (cat number 631294) would be suitable since they provide all necessary viral proteins and a transfection reagent. You can clone the flavivirus envelope into the pEmpty vector included with that kit or utilize your own vector. Alternatively, if you're looking for a reporter vector, cat number 631295 is also an option. This kit includes reporters along with the pEmpty vector for cloning purposes and is designed similarly to the 631294 kit. Although it’s marketed for SARS-CoV-2 envelope applications, it does not contain any COVID genes and should work well for integrating flavivirus envelopes as needed in your experiments.

recommend using total rna again for cdna synthesis to avoid rrna contamination?
Using total RNA for cDNA synthesis is indeed a recommended approach to minimize rRNA contamination. Total RNA typically provides a more comprehensive representation of the transcriptome and reduces the risk of losing smaller mRNA fragments during size-fractionation processes. Additionally, it allows for better control over potential degradation issues that can arise with isolated mRNA or fragmented samples. It's also beneficial to check the RIN value and 28S/18S ratio prior to use, as these metrics will help ensure that your RNA is of high quality and free from impurities, which ultimately supports successful cDNA synthesis.

what is the maximum insert size I can achieve using infusion with split primers for my specific application?
The maximum insert size you can achieve using In-Fusion with split primers will depend on the project mode you choose. If you're in Mutagenesis Project mode, it allows for a maximum insertion of 65 bp. However, if you opt for Cloning Project mode, larger inserts are possible; we've successfully worked with inserts up to 35 kb in that context. For your specific application and based on your requirements, using Cloning Project mode would be advisable if you're looking to insert a more significant sequence.

can i still recover rna samples after not taking supernatant from beads during ribosomal depletion?
If you did not remove the supernatant from the beads during ribosomal depletion, it may affect the quality and quantity of your RNA samples. However, there is still a possibility to recover some RNA. You can try to resuspend the beads in an appropriate buffer and perform additional washes or extraction steps to see if you can isolate any remaining RNA. It's important to assess the integrity and purity of the recovered RNA afterward, as these factors will influence your downstream applications.

Can I change my order to just five vials instead of packs?
To change your order to just five vials instead of packs, it is best to directly contact Customer Support. They will be able to assist you with modifying your order and provide the necessary instructions. You can reach them by phone or email during their business hours for prompt assistance.

can you confirm the percent recovery method used for validating the viral concentration?
The percent recovery method for validating viral concentration typically involves comparing the viral titer before and after purification. To accurately estimate recovery, it's essential to know the initial viral titer. A high recovery percentage, ideally around 90%, is expected when using a silica membrane due to its strong binding capacity. It's also recommended to utilize a specific titration kit that aligns with your virus type and ensure compatibility with your primers for accurate results.

which smart-seq mrna kit should i use for obtaining transcripts from single cells?
For obtaining transcripts from single cells, the SMART-Seq mRNA Single Cell Kit is the recommended option. This kit is specifically designed for single-cell analysis and offers higher sensitivity compared to the standard SMART-Seq mRNA kit. It utilizes enhanced Reverse Transcriptase (SMARTScribe II) and allows for compatibility with a range of cell numbers, from one to one thousand intact whole cells. The technical note highlights its unprecedented sensitivity, making it ideal for detailed transcriptomic studies at the single-cell level.

what kind of elements are crucial for effective packaging in the system you mentioned?
Effective packaging in HIV-1 based lentiviral systems typically requires several crucial elements, including the following: 1. **Packaging Plasmids**: These include plasmids that encode essential viral proteins such as Gag, Pol, and Env. The Gag protein is particularly important for forming the viral core. 2. **Transfer Vector**: This vector contains the gene of interest along with necessary regulatory elements like promoters and enhancers to ensure proper expression once inside target cells. 3. **Envelope Protein**: Choosing an appropriate envelope protein is vital for determining host range and enhancing infection efficiency. Commonly used envelopes include VSV-G (Vesicular Stomatitis Virus Glycoprotein) which provides broad tropism. 4. **Self-Inactivating (SIN) LTRs**: These are modified long terminal repeats that help prevent re-infection by ensuring that after integration into a host genome, the viral genes cannot produce new virus particles from infected cells. 5. **Helper Proteins**: In some systems, additional helper proteins may be included to support efficient assembly and release of the lentivirus particles. These components work together to create a robust system capable of producing high-titer lentiviral vectors suitable for various applications in research and therapy.

how do I charge sales tax for my order in michigan?
To charge sales tax for your order in Michigan, you should first determine the appropriate sales tax rate based on the type of product being sold. In Michigan, the general state sales tax rate is 6%. If you're selling taxable goods or services, you must collect this amount from customers at the time of sale. Ensure that you register with the Michigan Department of Treasury to obtain a sales tax license if required. It's also important to keep accurate records of all transactions and remit collected taxes as per state guidelines. For specific questions regarding compliance or unique situations, consulting with a tax professional may be beneficial.

Greet
Hi $$$visitor_first_name$$$. I'm $$$Operator_First_Name$$$, Thank you for your interest in Takara products. How may I assist you today?

what sequencing depth and kit do you suggest for smart-seq to effectively detect genes and transcripts?
For effective gene and transcript detection using SMART-Seq® 3' DE, it's important to consider both sequencing depth and the specific kit used. The recommended approach is to utilize a high sensitivity method, as demonstrated by the end capture technique employed in this kit. A typical setup involves ensuring sufficient input quantity; ideally, individual single-cell libraries should be sequenced at depths that can provide reliable coverage across expressed genes. In practice, variability in cDNA yield from different cells means that read counts per cell may vary significantly—ranging from approximately 0.2 million to 5.0 million reads as observed with K562 cells in pooled samples. Thus, aiming for a higher sequencing depth will enhance the likelihood of accurately detecting differential expression among genes while taking advantage of the capabilities provided by the SMART-Seq® v4 technology. Consider optimizing your experimental design based on these parameters alongside pilot studies to determine the ideal conditions specific to your samples and research objectives.

why haven't i received my password?
It looks like your password is provided by Storehelpers, and they are currently not available for chat support. The best option would be to contact them directly via email for assistance with your password inquiry.

need know construct design acceptable
To determine if your construct design is acceptable, you can use the Primer Design Tool on the Takara website. By entering your sequence information there, you'll be able to generate a primer design file. If you'd like assistance with this process, please click on "Report Issues/Ask Question" at the bottom of the tool's page after you've entered your data. Fill out the form and submit it; this will send your file directly to Takara for analysis. The team will review your design and get back to you via email with feedback. Let me know if there's anything else I can assist you with!

Looking coa?
The Certificate of Analysis (CoA) can be found at this link: https://catalog.takara-bio.co.jp/PDFS/T100A_AO4G003_y_COA.pdf. If you need any further assistance or have additional questions, feel free to ask!

should I be concerned about the efficacy of reagents if they're kept on ice?
Keeping reagents on ice during use is generally acceptable and should not significantly affect their efficacy. It's important to avoid letting them reach room temperature, especially for sensitive components like polymerases. Proper storage conditions help maintain the activity of these reagents, so as long as they are kept on ice and handled carefully, you shouldn't be overly concerned about their effectiveness.

Greet
Hi $$$visitor_first_name$$$. I'm $$$Operator_First_Name$$$, Thank you for your interest in Takara products. How may I help you today?

hi placed order not received confirmation email can you check my order status?
It seems there may have been an issue with your order confirmation email. I recommend reaching out to customer service for assistance in checking the status of your order. If you provide them with any identifying information related to your order, they should be able to help you more effectively. Since their representatives are currently busy, it might take a little time for them to get back to you. You can also ask if they could follow up via email once they're available. Thank you for your patience!

what's the best approach to assess DNA contamination in my samples using intergenic regions for bacterial transcripts?
To assess DNA contamination in your samples, particularly when focusing on intergenic regions for bacterial transcripts, consider the following approach: 1. **Analyze Library Composition**: Begin by examining the distribution of reads within your library. A high percentage of intergenic reads may indicate potential genomic DNA contamination. 2. **Inspect Size Distribution**: Check if there is a peak corresponding to high molecular weight DNA alongside the expected library peaks in your size distribution analysis. This could also suggest contamination with genomic DNA. 3. **Perform Mapping and BLAST Analysis**: Conduct strand-agnostic mapping of your reads to identify their origin more accurately, especially if you suspect contamination. Additionally, perform a BLAST search against bacterial databases to see if any significant matches emerge from intergenic regions. 4. **Quantitative Assessment**: If possible, quantify the proportion of intergenic versus coding region reads to get a clearer picture of contamination levels. 5. **Consider Degradation Factors**: Be aware that degraded DNA can still incorporate into libraries; hence maintaining an eye on overall sample quality during preparation is essential. By combining these strategies, you'll gain better insights into whether there's significant DNA contamination affecting your RNA-seq analysis related to bacterial transcripts.

Greet
Hi $$$visitor_first_name$$$. I'm $$$Operator_First_Name$$$, I hope you are well. How may I help you today?

Greetings
Hi $$$visitor_first_name$$$. I'm $$$Operator_First_Name$$$, I hope you are well. How may I help you today?

tell exactly how cloning takar race works
The SMARTer RACE kit from Takara Bio utilizes the In-Fusion technology for cloning. Here's how it works: 1. **Template Preparation**: The kit includes a Template Switching Oligo that enhances cDNA synthesis, allowing for the creation of full-length cDNA from RNA templates. 2. **PCR Amplification**: After synthesizing the cDNA, PCR amplification is performed to generate the desired product. 3. **Directional Cloning**: The resulting PCR product contains 15-mer overlaps with a linearized vector provided in the kit. This overlap facilitates directional cloning, which ensures that the insert is inserted into the vector in a specific orientation. 4. **In-Fusion Assembly**: The linearized vector and PCR product are mixed together with In-Fusion enzyme in a single reaction step. This enzyme promotes homologous recombination at the overlapping regions. 5. **Transformation**: Finally, this mixture is transformed into E.coli cells to propagate and select for clones containing your insert. This method provides an efficient and reliable way to clone DNA fragments directionally into vectors compared to non-directional methods such as TA-cloning.

why is my protein yield so low in the expression vector?
There could be several reasons for low protein yield in your expression vector. It might relate to the specific cell type you're using, as different cells can have varying efficiencies in expressing proteins from certain vectors. Additionally, the transfection reagent and method you’re employing may not be optimal for your particular system. Consider checking the health of your cells during the experiment, as unhealthy cells can lead to reduced expression levels. Also, ensure that you've properly designed your construct with appropriate restriction sites and regulatory elements. It’s worth noting that time points post-transfection can affect yields; if possible, testing longer incubation times might provide better results. If you've consistently observed unexpectedly low expression despite these factors being accounted for, reaching out to colleagues or experts familiar with this specific vector or strain could provide further insights into potential issues. Continuing communication with technical support teams or R&D departments might uncover nuances within their constructs that could explain the disappointing results you are experiencing.

am looking for an extraction kit that works with the king fisher automated extraction analyzer, can you help?
The NucleoMag® Pathogen kit is a suitable option for cell-free DNA purification and works seamlessly with the KingFisher automated extraction analyzer. It can handle various sample types, including less than 200 µl of whole blood, serum, plasma, feces, and swab wash solutions, as well as up to 30 mg of tissue. For more detailed information on its usage and capabilities, you may want to refer to the user manual available online. If you have further questions or need assistance with anything else related to this kit or extraction processes in general, feel free to ask!

am looking for extraction kit for urine, stool, wound, and nail clinical samples using king fisher automated extraction analyzer?
The NucleoMag® Pathogen kit you mentioned is primarily designed for the purification of viral DNA and RNA from various sample types, including whole blood, serum, plasma, tissue samples, feces, and swab wash solutions. While it works well with KingFisher systems and can handle a range of volumes and materials effectively, it may not be specifically optimized for urine or nail clinical samples. For your needs regarding extraction from urine, stool, wound specimens, and particularly nails—which can indeed be challenging—it's advisable to look for kits explicitly tailored to those sample types. Consider checking the product specifications or contacting the manufacturer directly to confirm compatibility with all desired sample matrices. Additionally, they may offer other kits better suited for extracting DNA/RNA from urine or tougher materials like nails.

how can i update my ach information and get the correct ap phone number to resolve my issue?
To update your ACH information, it's best to reach out directly to the Accounts Receivable department. They will be able to assist you with the necessary steps. Additionally, for any questions regarding accounts payable or related issues, they can provide you with the correct contact number. If you have any other inquiries or need further assistance, feel free to ask!

how do nucleospin columns efficiently bind plasmid dna?
NucleoSpin columns efficiently bind plasmid DNA through a process that leverages the properties of silica membranes. Under high salt conditions, the DNA molecules interact with the silica, allowing them to adhere to the membrane while contaminants remain in solution. This binding is facilitated by specific buffer compositions that create an optimal environment for the interaction between the DNA and silica. After washing steps to remove impurities, elution buffers are used to release the purified plasmid DNA from the column, ensuring a high-quality yield suitable for further applications.

mean rna sample mirna?
The conversation indicates that there is a cutoff size of 200 nucleotides for RNA fragments, meaning any fragments shorter than this will not be retained in the column. For isolating miRNA specifically, a special kit called NucleoSpin® miRNA (740971.50) is recommended. This kit is designed to effectively isolate miRNAs from samples, ensuring optimal recovery and purity for downstream applications. If further information or support is needed regarding RNA or miRNA isolation, reaching out to technical support can provide additional assistance.

can you confirm if the bigger vials for the hep differentiation kit have been sent?
It appears that the larger vials for the hep differentiation kit were indeed sent, as noted in your message about sending Y30016 with a list of items. However, it would be best to wait for confirmation from orders regarding this shipment since you mentioned submitting the form recently and haven't received an update yet. Keep an eye out for that confirmation to ensure everything is on track.

centrifuged monday?
It seems like the centrifuge process is expected to take place on Monday. If you need any further details or clarification about the schedule or procedures, feel free to ask!

Can you provide more details about your return policy?
The return policy details can vary based on the specific product and circumstances of the purchase. It's best to consult our customer service team for accurate information tailored to your situation. They will be able to assist you with any questions regarding returns, including timeframes and conditions. If you need further assistance, please let me know!

what steps can we take to improve protein yield and minimize contaminants during purification with talon resin?
To improve protein yield and minimize contaminants during purification with Talon resin, consider the following steps: 1. **Evaluate Sample Nature**: Confirm if the sample is bacterial or mammalian, as this can influence binding characteristics. 2. **Check Binding Capacity**: Ensure that your protein concentration does not exceed the binding capacity of 15-18 mg/ml of resin. If it does, you may need to scale back your input. 3. **Assess Tag Exposure**: Conduct a pilot study under denaturing conditions to determine if the His-tag on your protein is properly exposed for binding. Improved yield in these conditions would indicate an issue with tag exposure under native conditions. 4. **Optimize Wash Buffer Composition**: Incorporate imidazole into your wash buffer to help reduce non-specifically bound proteins while retaining your target protein. A concentration of 10-20 mM imidazole could be effective. 5. **Consider Alternative Buffers**: While ammonium or sodium acetate buffers are sometimes used with nickel resins, carefully evaluate their compatibility with Talon resin based on specific experiences or literature references. 6. **Review Elution Strategy**: Ensure that elution steps are optimized for maximum recovery without compromising purity by adjusting imidazole concentrations accordingly between washes and elutions. 7. **Monitor Contaminants**: Analyze any contaminants present in final products closely; understanding their nature might provide insights into further adjustments needed during purification processes. By implementing these strategies, you can enhance both the yield and purity of your purified protein using Talon resin.

what's the difference between the lp kits and the ones that don't say lp?
The primary difference between the LP (Library Prep) kits and those that do not specify "LP" lies in their intended use and components. The SMART-Seq® mRNA LP kit is specifically designed for converting full-length double-stranded cDNA into an Illumina sequencing library, utilizing Unique Dual Indexing primers that must be purchased separately. In contrast, the kits without the "LP" designation typically focus on other aspects of RNA sequencing preparation but may not include these specific library preparation capabilities or additional indexing features. So, if you're looking to perform library prep for sequencing, you'll want to choose a kit labeled as "LP," while other kits might serve different roles in your workflow.

what's one good alternative for thermofisher hifi library prep?
One good alternative for Thermo Fisher's HiFi library prep is the PrimeSTAR® GXL DNA Polymerase from Takara Bio. It has been validated for use in PacBio sequencing and can handle long-range PCR applications, making it suitable for generating high-quality libraries. Additionally, it comes in both standalone and 2X premix formats, providing flexibility based on your workflow needs. Another option is the PrimeSTAR® LongSeq DNA Polymerase, which supports even longer amplicons of up to 50 kb and is also compatible with PacBio technology. Both options are effective alternatives for library preparation in various sequencing applications.

can you help me with that?
Yes, I can help with that. I'll forward the information to your territory manager, who will get back to you shortly. If you have any other questions or need further assistance, feel free to let me know!

What is the reason for my call?
The reason for your call could be to seek assistance, ask questions, or discuss a specific topic of interest. It's important to clarify what you need help with so that the conversation can be as productive and helpful as possible. What specific information or support are you looking for today?

how can i analyze the results of my bcr sequencing run?
To analyze the results of your BCR sequencing run, start by using appropriate bioinformatics software that is compatible with your data type. You may consider tools like IGV (Integrative Genomics Viewer) or other specialized software designed for BCR analysis, which can help visualize and interpret the sequencing data. It's essential to assess the quality of your library first. If you have generated Bioanalyzer traces, review them for size distribution and concentration to ensure adequate library preparation before proceeding with analysis. Once you've analyzed the raw sequencing data, look at key metrics such as read count, quality scores, and any alignment statistics provided in sequencer-generated reports. This information will be crucial in understanding whether there are issues affecting your results. If you're experiencing low read counts or other specific concerns regarding your sequencing output, sharing those details along with any relevant reports or BA traces would be beneficial for more targeted troubleshooting assistance. Feel free to reach out via email if you need further support or advice on interpreting specific aspects of your results.

why gostix value concentrating similar concentrating
The similarity in the GV (golden value) readings before and after concentration can often be attributed to a few key factors. When you concentrate a viral sample, the goal is to enhance the signal associated with virus particles, particularly virus-associated p24. However, if your dilution factor post-concentration effectively offsets any increase in concentration of the virus, it could result in similar GV values. In this scenario, if you used a 1:1 dilution for your pre-concentrated sample and then conducted a 1:10 dilution on your concentrated sample but obtained comparable GV outputs of 500 for both samples, this suggests that while concentrating may have increased the amount of infectious viral material present, the subsequent dilution minimized its apparent effect in terms of signal readout. Additionally, successful precipitation would indicate effective recovery during concentration; however, assay variability may also play a role. If both assays yield consistent percentages of infection despite these changes in preparation methods and dilutions, it reflects good consistency across experiments. Overall, it's essential to consider all variables involved—such as time spent precipitating and how well each step was optimized—to fully understand why GV values appear similar despite differences in processing steps.

what could be causing the unusually large viral pellet and how can I successfully resuspend it?
The unusually large viral pellet could be attributed to an increase in protein concentration, possibly due to changes in the health of packaging cells at the time of harvest. Although there haven't been any procedural changes, variations in cell viability or other environmental factors might lead to a higher amount of protein being present. To successfully resuspend the pellet, consider using a larger initial volume for resuspension. Starting with 440 µL instead of 220 µL may help dissolve the pellet more effectively. You can gradually add small volumes of media until full dissolution is achieved. Additionally, utilizing a concentrator with a 100 kDa cut-off unit will allow you to later reduce your volume back down if needed. If you're concerned about storage and stability, remember that using the concentrator allows for short-term storage at 4°C without negatively impacting virus integrity. It's also advisable not to remove the supernatant while resuspending since it may contain valuable components that contribute to your solution's effectiveness.

need pay lisence list price?
It seems that for non-profit organizations, the license may be available for free, while for-profit entities would typically need to obtain a paid license. For precise details and any potential costs associated with obtaining a license, it's best to contact the licensing group directly. They will provide the most accurate and up-to-date information regarding pricing and licensing terms.

am wondering if long digestion time could lead to different cloning efficiency or background issues?
Long digestion times can indeed affect cloning efficiency and background issues. Extended incubation may lead to partial digestion, where some of the vector remains uncut alongside fully linearized DNA. This can result in a higher number of colonies on plates due to the presence of intact vectors that allow for transformation. In addition, prolonged digestions might also increase nonspecific cleavage or degradation of the DNA, potentially impacting your insert's viability during ligation. To ensure optimal results, it's important to monitor digestion time carefully and validate complete digestion by running a gel analysis before proceeding with ligation or transformation. If you notice an excessive number of colonies after transformation, it could indicate incomplete digestion as well as other factors at play such as contamination or insufficient selection pressure.

how do i concentrate precipitate aav?
To concentrate AAV, you can use a concentrator designed for this purpose. While it's been reported that some users have successfully concentrated AAV using these methods, specific protocol modifications may be necessary depending on the system used and your particular conditions. If you're looking for both concentration and purification of AAV, consider utilizing dedicated AAV purification kits available in the market. These kits are specifically formulated to handle the unique requirements of AAV while ensuring effective concentration and purification from your media. Always refer to the manufacturer’s guidelines or user feedback for optimal results tailored to your setup.

can you please confirm the cancellation of my orders?
To confirm the cancellation of your orders, please reach out to the customer service team via email or phone. They will assist you with canceling one of the double orders you've placed. Currently, they are in a meeting until approximately 2 PM PDT and won't be available on chat or through technical support for this matter. If there's anything else you need help with, feel free to ask!

customer looking recommendations for best polymerase sensitive enough to use with low input purified gdna crude dna extracts
For low input purified genomic DNA (gDNA) from crude extracts, the TaKaRa Ex Premier™ DNA Polymerase is a good option; however, it's important to note that it may not be ideal for multiplexing applications. If you require high sensitivity, consider using Titanium Taq DNA Polymerase, which is highly sensitive and can work with as little as 10 ng of gDNA. Typically, input amounts range from 30 to 100 ng for standard applications, but if your samples are particularly sensitive or you're working with lower quantities, starting at 10 ng could yield good results. For multiplexing needs specifically, SuperPlex would also be suitable if you’re looking for optimized performance in those situations.

why is the search bar unavailable?
The search function on the website is currently down, which is causing issues for users trying to find specific information. However, the rest of the website remains accessible, and you can still browse through other sections to locate what you need. If there's anything else needed or if you'd like assistance in finding a particular document, feel free to ask!

how can i place an order for the smarter smrna-seq kit by illumina shipped to toronto?
To place an order for the Smarter smRNA-Seq kit by Illumina to be shipped to Toronto, you will need to follow these steps: 1. Visit the supplier's website or contact their sales team directly to select the product and confirm availability. 2. Ensure that you place your order before 2 PM Eastern Time on a weekday for it to be processed quickly; however, note that orders placed on Fridays will typically ship on Monday due to shipping policies. 3. If you're ordering from Canada, it's important to consider customs clearance options. Using express customs clearance can help avoid delays and ensure smooth delivery. 4. If possible, work with a broker who is familiar with dry ice shipments if your product requires it. Once you've completed these steps, you'll receive confirmation of your order and estimated shipping details.

why are the qc result peaks different from the manual?
Differences between QC result peaks and those in the manual can occur for several reasons. It may be due to variations in sample preparation, differences in instrument calibration, or even changes in the assay conditions. It's also possible that there are updates or revisions in the methodology that aren't reflected in the manual. To address this issue effectively, it would be beneficial to review your procedures against both the current protocols and any relevant documentation. If discrepancies persist, reaching out to technical support with specific data might help clarify these differences further.

how can we properly count the foci that look small and pushed together versus the larger foci?
To properly count the foci, it can be helpful to use images that clearly show both small and larger foci. By sending in these pictures, you can receive targeted feedback on how to differentiate between them. Annotating specific areas of interest in the images will also aid in discussions about counting methods. Additionally, involving colleagues with more experience might provide further insights into effective techniques for distinguishing and counting different sizes of foci accurately.

what steps do i need to follow to log off and resolve the software issue?
To log off and resolve the software issue, follow these steps: 1. Save any ongoing work to prevent data loss. 2. Close all open applications related to the software that is experiencing issues. 3. Log off from your user account by navigating to the appropriate option in the application or system settings. 4. If needed, restart your device to clear temporary files and refresh system resources. 5. After restarting, log back into the application and check if the issue persists. If you continue to experience problems, consider reaching out for further assistance via chat or email as mentioned during your team meeting.

why isn't the click link working?
There could be several reasons why the clickable link isn't working. It might be due to a broken URL, an issue with your internet connection, or restrictions placed by your browser or network settings. Additionally, if the webpage has been updated or moved since it was last accessed, that could also affect functionality. If troubleshooting doesn’t resolve the issue, consider reaching out via email for further assistance or checking back later to see if the problem persists.

access hv protocol provide customer fill form link?
It seems that the protocols and catalog numbers may not have been updated on the intranet. If customers require access to the HV protocol, it's important to provide them with a link or information about where they can find it. Additionally, if there are any changes in catalog numbers that haven't been communicated, be sure to inform customers so they have accurate details for their inquiries.

how can i place a long order?
To place a long order, it's best to start by reaching out to the company directly through their customer service or sales team. They can guide you through the process and provide any necessary forms or information specific to your needs. If you're an existing research or business account holder, mentioning that may help expedite your request. Make sure to clearly outline what products you need and in what quantities, as well as any other specifications. If needed, they will follow up with you for further details or confirmation.

what steps should i take to resolve the software issue and log off?
To resolve the software issue, you should first ensure that you're using the latest version of the application and check for any updates that may have been released. If there are specific instructions from support regarding changes made in the recent update, make sure to review those carefully. If it appears to be working now but requires some adjustment to the new view, take a moment to familiarize yourself with any updated features or layouts. For further assistance, consider reaching out to your IT support team via chat or email if issues persist. Once you've resolved your issue and feel comfortable with the new setup, you can log off by following these steps: save any work you may have open, close all applications related to this software, and then use the designated logout option typically found in your profile menu or settings tab within the application.

What products do you offer for cell concentrating and packaging reagents?
The conversation did not provide specific information about products for cell concentrating and packaging reagents. However, you can explore the resources available on the Takara Bio website, particularly in their learning center focused on viral transduction and adeno-associated virus (AAV) applications. This page includes links to various associated products that might meet your needs. If you have further questions after reviewing the materials, feel free to reach out again for more assistance.

hi felipe what are the patent details for the fluorescent protein plasmids listed on your website?
The product page for the AcGFP1 fluorescent protein plasmid indicates that it is not off patent, as there are no check marks under the license heading. This means a license is required for research use. For commercial applications, you will need to reach out to our licensing department for more information on obtaining the necessary rights. If you have any further questions or need assistance with anything else, feel free to ask!

looking for best options discounts on mrna delivery products while cross selling additional kits?
For mRNA delivery products, consider exploring our established range of options that have been trusted in the market for 15-20 years. While we currently do not have sample sizes available for these antibodies, you might find value in our green premixes like RR820ATB Green® Premix Ex Taq™ II (Tli RNase H Plus) and 639676-TB Green® Advantage® qPCR Premix. Both are excellent choices if you're also looking to enhance your qPCR capabilities. Regarding discounts, it's advisable to check for any ongoing promotions on bulk purchases or seasonal sales directly through our website or customer service. Additionally, bundling your order with complementary kits may provide further savings opportunities. If there’s a specific kit you’re interested in or if you need assistance finding suitable combinations, feel free to ask!

what is the highest fidelity product you offer that includes primestar gxl premix fast dye plus relative taq?
The highest fidelity product that includes PrimeSTAR GXL is the R045A-PrimeSTAR® Max DNA Polymerase. It offers a high fidelity rate with very low error rates, making it suitable for applications that require precise amplification. For more details on this polymerase and to explore its features, you can visit the product page on the manufacturer’s website. If you have any further questions or need assistance with your order, feel free to reach out to technical support or your territory manager.

What are the best options for reconstructing functional antibodies from sequencing data in smart-seq human BCR workflows using FFPE samples?
Reconstructing functional antibodies from sequencing data in SMART-Seq® Human BCR workflows using FFPE samples presents several challenges due to the degradation and chemical modifications often found in FFPE RNA. Here are some potential options to consider: 1. **Alternative Library Preparation Methods**: While current TBUSA immune profiling kits may not be compatible with FFPE RNA, exploring other library preparation methods designed specifically for degraded or low-quality RNA could be beneficial. Look for protocols that can accommodate truncated transcripts. 2. **Targeted Amplification**: Consider using targeted amplification strategies that focus on specific regions of interest within the BCR genes, which may help capture more relevant sequences despite the overall quality of the starting material. 3. **Use of UMI-Based Technologies**: If available, utilize unique molecular identifier (UMI) technologies that can enhance sensitivity and accuracy in quantifying BCR sequences even when starting with less-than-ideal RNA quality. 4. **Specialized Kits for FFPE Samples**: Research vendors who offer specialized kits or protocols tailored for working with FFPE-derived RNA, as these might include optimized reagents and methodologies designed to mitigate issues like truncation and chemical modifications. 5. **Bioinformatics Approaches**: Employ advanced bioinformatics tools to analyze sequencing data from partially degraded transcripts, allowing reconstruction of full-length antibody sequences based on existing sequence information. 6. **Collaboration with Specialized Labs**: Partnering with laboratories experienced in handling FFPE samples could provide access to expertise and resources necessary for successful antibody reconstruction from such challenging materials. Continuously monitoring advancements in this area is essential since new technologies or methodologies are being developed regularly that may improve outcomes when working with FFPE samples.

wondering if old rna cdna kits expired could be affecting qpcr results?
Using expired RNA or cDNA kits can definitely impact qPCR results. Expired kits may lead to reduced efficiency and sensitivity in your reactions, potentially resulting in unreliable data. It's important to ensure that all reagents are within their shelf-life for optimal performance. If you suspect that the kit you’re using is expired, it would be advisable to replace it with a fresh one before proceeding with your experiments. Additionally, always check the integrity and quantity of RNA prior to reverse transcription, as this can also influence results significantly.

why can't I detect qpcr when amplifying multiple preimplantation embryos?
There are several factors that could affect the detection of qPCR when amplifying multiple preimplantation embryos. One primary concern is the efficiency of lysis, especially considering the presence of the amniotic sac. If the embryos are not lysed effectively, RNA may remain intact and inaccessible for amplification. Using a freeze-and-thaw method on dry ice prior to adding lysis buffer can help facilitate better lysis by breaking down protective structures like the amniotic sac. It’s important to perform this step quickly and prepare your buffers in advance so that you can add them immediately after freezing. Additionally, ensure that you’re working with an appropriate volume of lysate for qPCR. The general rule is to use no more than 1/10th of your total reaction volume; for a 25 µl qPCR reaction, this means using no more than 2.5 µl of lysate. Excessive amounts can inhibit the reaction and lead to poor amplification results. Lastly, make sure your samples are well protected during processing to prevent RNA degradation, as compromised samples might also result in undetectable or low levels of amplification during qPCR analysis.

how can I improve the amplification results for multiple mouse preimplantation embryos using kit amplify?
To improve amplification results for multiple mouse preimplantation embryos using the kit, consider a few strategies. First, ensure that you effectively lyse the embryos before adding the lysis buffer. The presence of the amniotic sac can hinder efficient lysis, so applying a freeze-and-thaw method on dry ice prior to adding the lysis buffer may help break it open and enhance cell access. Additionally, confirm that you're working with an appropriate number of cells per embryo and optimize your PCR conditions based on those variables. If you encounter specific challenges or have further questions as you proceed, reaching out to technical support may provide tailored guidance for your situation.

how's your evening going?
The evening is going well, thank you for asking! It's a nice time to reflect and unwind after the day's activities. How about yours?

What is the best way to train my cat?
Training a cat can be a rewarding experience that enhances your bond. Here are some effective tips for training your cat: 1. **Positive Reinforcement**: Use treats, praise, or playtime as rewards when your cat follows commands or exhibits desired behaviors. 2. **Short Sessions**: Keep training sessions brief, around 5-10 minutes, to maintain your cat's attention and enthusiasm. 3. **Consistency**: Use the same cues and commands consistently so your cat can learn to associate specific words with actions. 4. **Patience is Key**: Cats may take longer than dogs to grasp new concepts, so be patient and understanding throughout the process. 5. **Start with Basic Commands**: Teach simple commands like "sit" or "come". Begin in a quiet environment free from distractions. 6. **Use Clicker Training**: A clicker can help mark the exact moment of a desired behavior, making it easier for your cat to understand what they did right. 7. **Make It Fun**: Incorporate play into training; use toys that encourage movement as part of learning activities. 8. **Respect Their Limits**: Not all cats will respond well to every type of training; it's important to recognize their individual personalities and preferences. By following these guidelines, you can create an enjoyable training experience for both you and your feline friend!

what's the best way to ensure proper virus collection and filtering in my lentiviral supernatant process?
To ensure proper virus collection and filtering in your lentiviral supernatant process, start by centrifuging the harvested supernatant to separate any cell debris. If you notice a significant amount of cell debris in the pellet, using a 100 kD Amicon filter can help purify your sample further. Filtration through a 0.45 μm filter is recommended for additional clarity and removing smaller particles, but it’s optional if you don’t have one available. If your target cells are sensitive, consider performing filtration after solubilizing the pellet to avoid damaging them. If the cells aren't very sensitive, you may skip this step without issue. Always assess your specific situation and adjust accordingly based on the sensitivity of your target cells and the presence of cell debris.

Where can I find a qPCR primer design tool online?
You can find several online tools for qPCR primer design at IDT. They offer a range of free tools under their SciTools Web Tools section, which includes: - **PrimerQuest Tool**: This tool allows you to generate customized designs specifically for qPCR assays and PCR primers. - **PrimeTime™ qPCR Assay Selection Tool**: Use this tool to select predesigned sequences targeting human, mouse, and rat genes. - **RealTime qPCR Design Tool**: This is useful for designing primers and probes across exon boundaries for gene targets in various species beyond just human, mouse, or rat. - **OligoAnalyzer Tool**: Analyze oligonucleotide properties such as melting temperature, potential secondary structures (like hairpins and dimers), and perform BLAST analysis directly from this platform. - **UNAFold Tool**: It helps analyze the secondary structure of oligonucleotides. These resources should help you get started with your primer design effectively.

What are my alternatives regarding this issue?
There are several alternatives you can consider regarding this issue. First, you might want to explore different solutions or approaches that could address your concerns more effectively. Additionally, discussing the matter with colleagues or seeking advice from professionals in the field may provide valuable insights and options. If applicable, reviewing relevant policies or guidelines could also help clarify your choices. Finally, don't hesitate to reach out for further assistance if needed; collaboration often leads to better outcomes.

shall send plasmid sequence primers?
It would be helpful to send the plasmid sequence along with the primers. This will allow for a more thorough analysis of the GC content and any potential issues with primer specificity. Having this information can assist in redesigning primers that are better suited for your amplification needs, especially considering factors like Tm values and binding sites.

can you provide a quotation including the gel tray?
Certainly! The quotation will include the electrophoresis unit and a gel maker set, which consists of 4 small gel trays, 2 large gel trays, 2 large combs, and 1 large gel maker stand. Please confirm if this is acceptable or if you need any additional information.

where can I find Seles products and their resellers in Montreal, Canada?
Seles products may not be directly available through a distributor in Canada, but you can consider using third-party distributors like VWR to find and order these products. For specific inquiries or to place an order, contacting customer service via the provided phone number or email may also be helpful.

can you provide guidance on a hybridization-based protocol for TCR profiling in mouse DNA?
For hybridization-based TCR profiling in mouse DNA, consider the following key steps: 1. **Sample Preparation**: Start by isolating high-quality genomic DNA from mouse tissues or cells. Ensure that the extraction method preserves the integrity of TCR genes. 2. **Design Probes**: Design specific oligonucleotide probes that target various regions within the TCR gene segments (e.g., V, D, J). The probes should be complementary to known sequences and labeled for detection. 3. **Hybridization Conditions**: Optimize hybridization conditions such as temperature and time based on probe melting temperatures and specificity requirements. Use a controlled environment to minimize non-specific binding. 4. **Detection Method**: Choose an appropriate detection method post-hybridization, which can include fluorescence or chemiluminescence depending on your probes' labeling strategy. Confirm signal strength and specificity through control experiments. 5. **Data Analysis**: Utilize bioinformatics tools for analyzing hybridized signals to determine TCR diversity and clonality across samples effectively. Always validate your protocol with known standards before proceeding with experimental samples to ensure accuracy in results.

when will the full length cDNA smart-seq kit be available for order online?
The full-length cDNA SMART-seq kit compatible with Oxford Nanopore is expected to be released on March 11, 2025. On the release date, the protocol will also be available online on the Takara Bio USA website. Before ordering, it's recommended to read the User Manual for details on included reagents and any additional materials that may need to be purchased separately.

why is the app not working for band quantification data?
The app may not be functioning properly for band quantification data due to a few potential reasons. It could be experiencing temporary downtime, as noted in your earlier message about it being down yesterday. Additionally, if multiple devices across different operating systems are having issues, this might indicate a broader compatibility problem or software bug. Moreover, it's important to consider that the accuracy of the band quantification can diminish over time due to factors like evaporation; results are best collected within 10 to 15 minutes after application. If problems persist despite restarting your device and checking other settings, notifying the app host would be a good step toward resolving any underlying issues.

What calculations do I need to perform for the gv value after receiving the dilution baculovirus?
To calculate the gv value after receiving the dilution of baculovirus, you need to consider whether you've inputted the dilution into your application. If you have entered the dilution correctly, it will automatically adjust and provide you with an accurate result. However, if you did not include this information in your calculations, you'll need to multiply the answer obtained by 10 to account for the dilution factor. This adjustment ensures that you're reflecting the true concentration of baculovirus in your final calculation.

which competent cells should I use with the in-fusion cloning kit?
For the In-Fusion cloning kit, it's recommended to use competent cells that are specifically designed for high-efficiency transformation. Common choices include commercially available competent cells such as NEB 5-alpha or One Shot™ TOP10 cells. These options are compatible with the In-Fusion system and should work well for your cloning needs.

can you provide more information on how to improve transduction efficiency for my specific application?
To improve transduction efficiency for your specific application, consider the following strategies: 1. **Use Transduction Enhancers**: Incorporating enhancers like RetroNectin can facilitate better binding of viral particles to target cells, especially in hematopoietic cell transduction. 2. **Optimize Viral Vector Concentration**: Experimenting with different concentrations of your retroviral vector may help identify the optimal amount needed for effective transduction without causing cytotoxicity. 3. **Increase Multiplicity of Infection (MOI)**: Increasing the MOI can enhance the likelihood that each target cell will be successfully infected by a viral particle. 4. **Cell Cycle Considerations**: Since retroviral vectors primarily transduce dividing cells, ensuring that your target cells are in an active division phase can yield better results. 5. **Pre-treatment Protocols**: Depending on your cell type, pre-treating them with agents such as cytokines or growth factors could stimulate proliferation and increase susceptibility to infection. 6. **Transfection Methods**: If using plasmid vectors instead of viral ones, you might explore various methods like electroporation or lipofection to maximize uptake into difficult-to-transfect cells. 7. **Review Compatibility with Cell Type**: Ensure that the chosen vector is compatible with the specific characteristics and requirements of your target cells for optimal expression levels post-transduction. For detailed guidance tailored to your needs, consulting technical notes from manufacturers or engaging directly with their support teams can provide valuable insights based on current best practices in gene delivery systems.

maybe can you provide more information about the plasmid and how it works as a vector?
The plasmid vector, specifically the pCMV-DsRed-Express2 Vector, is designed to enable the expression of the DsRed-Express2 protein in mammalian cells. This vector contains a strong cytomegalovirus (CMV) promoter that drives high levels of gene expression. The DsRed-Express2 protein itself is known for its minimal cytotoxicity and high solubility, making it suitable for various applications in cell biology. When transfected into target cells, the plasmid allows for stable integration and expression of the DsRed-Express2 fluorescent marker, enabling researchers to visualize cellular processes through fluorescence microscopy. The strong and stable expression provided by this vector can be particularly advantageous in experiments requiring long-term observation or tracking of cells. If you have any specific questions about applications or features, feel free to ask!

what are the best cells for propagating large plasmids with better stability?
For propagating large plasmids with better stability, HST08 cells are an excellent choice. They have a mostly intact DNA repair machinery, which helps maintain the integrity of larger plasmids during amplification. In contrast, standard cloning strains often possess defective DNA repair mechanisms that can lead to instability in large plasmids. Therefore, using a strain like HST08 is recommended for optimal results with large constructs.

is emerlandamp pcr master mix compatible with sequencing using green dye?
The compatibility of the EmeraldAmp PCR Master Mix with sequencing when using a green dye depends on several factors, including the specific absorbance characteristics of both the master mix dyes and the sequencing system. While some users have successfully sequenced samples containing loading dyes without issues, it's important to consider that certain dyes may cause interference during sequencing due to their absorbance properties. If your master mix contains yellow and blue dyes, there might be potential for interference if those wavelengths overlap with the detection range used in your sequencing process. However, cleaning up your PCR product before sequencing can help mitigate any possible effects from these dyes. If you're concerned about compatibility or interference, it’s advisable to perform a cleanup step or consult additional resources or protocols related to your specific application.

what's the status of my order?
The conversation provided does not contain any specific information regarding the status of your order. To obtain updates on your order, it's best to check directly with the customer service team or through the appropriate online platform where you placed your order. If you have a tracking number or order confirmation, that can also be helpful in getting detailed information about its status.

what do you think about this product?
The product in question seems to have a specific application for eukaryotic organisms due to its use of oligo dT priming. This targeted approach suggests that it may be particularly effective for studies involving eukaryotic RNA, making it a valuable tool for researchers in related fields. Its specialized nature could enhance the accuracy and efficiency of experiments focused on eukaryotic gene expression or other molecular biology applications. Overall, it appears to be a well-suited product for those working with eukaryotes.

why is lenti go stix app crashing when I try to enter sample names?
It seems that the Lenti Go Stix app may be experiencing technical issues, especially if it’s not functioning properly on your device. There could be various reasons for the crashes when entering sample names, such as compatibility problems with your specific device or operating system version, firewall settings interfering with app performance, or even recent updates affecting functionality. To troubleshoot this issue, you might want to check for any available updates for the app and ensure that you're using a stable internet connection—try switching between Wi-Fi and cellular data. If it has worked previously on your device without issues, there may currently be a temporary glitch affecting its operation. Given that there were website issues reported earlier in the day and this is the first complaint about the app crashing under these circumstances, it's possible there's an underlying problem being addressed by the developers. Reporting your experience can help them identify and resolve any bugs more efficiently. Thank you for your understanding while we work towards a solution!

how do i troubleshoot issues with gravity flow?
To troubleshoot issues with gravity flow in a column or liquid chromatography system, consider the following steps: 1. **Check Flow Rate**: Ensure that the flow rate is consistent and within the expected range. If it's too slow, it may indicate blockages or improper packing. 2. **Inspect Column Packing**: Make sure the column is packed evenly without voids or cracks. Poorly packed columns can lead to irregular flow patterns. 3. **Examine Connections and Tubing**: Look for any leaks, kinks, or obstructions in tubing and connections that could impede flow. 4. **Assess Mobile Phase Composition**: Verify that the mobile phase is prepared correctly and at an appropriate viscosity for optimal flow. 5. **Air Bubbles**: Check for air bubbles trapped in the system which can disrupt gravity flow. If present, carefully tap on tubing to release them. 6. **Column Height Adjustment**: Adjusting the height of your setup may help improve gravitational pull if you're experiencing very slow flows due to elevation differences. 7. **Temperature Control**: Ensure that temperature conditions are stable as fluctuations might affect fluid properties and thus alter flow rates. 8. **Visual Inspection During Operation**: Monitor changes during operation; unexpected variations can give clues about underlying issues requiring attention. Following these troubleshooting tips should help identify and resolve common problems associated with gravity-flow systems effectively.

can you confirm if the guide-it guide sgrna vitro transcription kit can be used for a portion of the workflow in sgRNA knockdown for bacteria?
The Guide-It sgRNA In Vitro Transcription Kit can indeed be utilized for generating sgRNA, which is a crucial part of the workflow for sgRNA knockdown in bacteria. While we do not have specific protocols or kits designed exclusively for gene editing in bacteria, you can combine the synthesized sgRNA with our recombinant Cas9 protein to create a ribonucleotide complex that may then be electroporated into bacterial cells. Though this approach has not been tested by us, there are likely relevant protocols available in scientific literature that could guide you through the process.

What is the cogent ngs analysis pipeline?
The Cogent NGS analysis pipeline is a comprehensive tool designed for the processing and analysis of next-generation sequencing (NGS) data. It typically includes several key steps such as quality control, alignment to reference genomes, variant calling, and annotation. The pipeline may also offer additional features like visualization tools and integration with databases for further insights into genomic data. For specific inquiries about its functionalities or technical aspects, it’s recommended to reach out to bioinformatics specialists who can provide detailed information tailored to your needs. Please consider sending an email outlining your questions for more precise support.

whats going on with my order?
It appears that there have been some delays with FedEx due to a winter storm in the Midwest, which may have affected your order. Unfortunately, it seems that your order was not received on our end. To resolve this, please resend your purchase order to the designated email address and include technical support in the correspondence for additional support. The latest record we have shows an order date of February 7, 2025. If you need further assistance or have any other questions, feel free to ask!

study data transduction efficiency vcn using retronectin please share
It sounds like you're interested in studying the correlation between transduction efficiency and vector copy number (VCN) when using retronectin. While there may not be direct evidence of retronectin modifying viral genomes, its role in enhancing transduction is well-established. To investigate this further, it would be beneficial to look into published studies or data that specifically examine VCN in conjunction with retronectin treatments. If you have access to relevant databases or publications, they might provide valuable insights. Additionally, considering factors such as how transduction efficiency was tested—like using a fluorescent reporter on the same vector—could help clarify results and ensure accurate interpretation. It's also important to explore methods for minimizing interference from broken viral genomes during your analysis. If there’s specific data you need or if you’d like assistance locating relevant studies, feel free to reach out once you've had some time to dig through your resources!

what can you help me with regarding my purchase?
I can assist you in checking the details of your purchase, including verifying any lot numbers associated with the items. If that information isn't available through our Orders system, I can coordinate with our QA department to gather additional insights. Please feel free to reach out if you have any specific questions or need further assistance regarding your order.

what key features should i consider when comparing the two kits for the head-to-head evaluation?
When comparing the SuperScript™ IV VILO™ Master Mix and PrimeScript™ RT Master Mix (Perfect Real Time) for a head-to-head evaluation, consider the following key features: 1. **Temperature Range**: SuperScript III operates at higher temperatures (50-55°C), which can help with secondary structure resolution but may risk RNA degradation. In contrast, PrimeScript RT works effectively at 42°C, which is gentler on RNA stability, especially beneficial for GC-rich templates. 2. **RNA Input Range**: Assess the recommended input RNA amounts for each kit. SuperScript IV VILO supports a broader range for TaqMan applications (1 pg to 2.5 µg) compared to PrimeScript's range of 2 pg to 2 μg in a standard reaction volume of 20 µL. 3. **Performance with Different Templates**: Consider how each enzyme performs with various types of templates, particularly GC-rich sequences where PrimeScript shows strong strand displacement activity. 4. **Sensitivity and Reproducibility**: Evaluate data or publications that demonstrate the sensitivity and reproducibility of cDNA synthesis from both kits under different conditions. 5. **Application Compatibility**: Identify if one master mix has advantages over another in specific applications like RT-qPCR using SYBR versus TaqMan assays due to differences in performance metrics highlighted in studies. 6. **Ease of Use and Protocol Efficiency**: Look into whether either kit offers simplified protocols or faster turnaround times that could benefit your workflow. By focusing on these aspects, you can make an informed decision based on your experimental needs and priorities.

what is the best method for quantifying aav genomic dna using the great takara kit?
The best method for quantifying AAV genomic DNA using the Takara kit involves a combination of cell permeabilization to release cytoplasmic contents without lysing the cells, followed by the use of Cryonase to degrade free nucleic acids. This approach helps in obtaining accurate measurements of AAV genomic DNA by minimizing contamination from extraneous nucleic acids. After preparing your samples appropriately, you can proceed with qPCR using the specific protocols provided in the Takara kit for optimal results.

what is the shelf life of protein once manufactured?
The shelf life of protein products can vary depending on the type and how they are processed, but typically, most protein powders and supplements have a shelf life of about 1 to 2 years from the date of manufacture when stored properly. It's important to check the expiration date on each specific lot for accurate information. Proper storage in a cool, dry place can help maintain quality throughout its shelf life.

why low yield from transfection compared to expectations?
The low yield from transfection compared to expectations could be due to several factors. One possibility is that the incubation time may have been longer than necessary, which can lead to a decrease in titer. Additionally, if the concentrator used for virus stabilization had not been mixed well or experienced evaporation, it might result in reduced performance and lower yields. It's also important to consider the specific conditions of your experiment, such as temperature during the spin and any changes in materials or reagents being used. If this is the first use of a particular lot of concentrator, it's worth checking for any inconsistencies as well. Overall, assessing these variables should help identify potential reasons for the unexpected results.

how can i convert xls files to csv and resolve errors encountered while following the manual?
To convert XLS files to CSV format, you can use various methods depending on the tools available to you. Here are a few options: 1. **Using Microsoft Excel**: - Open your XLS file in Microsoft Excel. - Click on "File," then select "Save As." - In the dialog box, choose "CSV (Comma delimited) (*.csv)" from the dropdown menu for file formats and save. 2. **Using Google Sheets**: - Upload your XLS file to Google Drive. - Open it with Google Sheets. - Click on "File," then select "Download" and choose “Comma-separated values (.csv).” 3. **Using Online Converters**: - There are several online conversion tools where you can upload an XLS file and download it as a CSV without needing software installed. Regarding errors encountered while following the Cogent NGS Immune Profiler User Manual, it's best to consult directly with Bioinformatics specialists for tailored assistance. They will be able to provide specific guidance based on the issues you're facing. If you'd like me to forward this inquiry for further support, please let me know!

what options do you have for takara products that perform better in the laboratory?
For enhanced laboratory performance, Takara offers a variety of products tailored for specific applications. The QIAcard FTA Elute Buffer is designed specifically for improved elution of nucleic acids from biological samples, making it suitable for sequencing and various PCR applications. Additionally, the T9111-Tris-EDTA (TE) Buffer 10X Powder allows users to prepare a customized TE buffer with precise concentrations by reconstituting it in distilled or deionized water. If you're looking for more options that might perform better in your specific experiments, consider exploring other specialized buffers and kits from Takara that are optimized for different types of assays, such as qPCR or STR analysis. These can provide enhanced sensitivity and specificity depending on your research needs. If you have particular application requirements in mind, further recommendations can be provided based on those specifics.

storage shipping temperature ones?
The storage and shipping temperatures for the products mentioned are as follows: - Product 632637 should be stored at -20°C and shipped with dry ice. - Product 632638 can be stored at room temperature and shipped at room temperature. Both products are components of product 632635. If you need further assistance or have more questions, feel free to ask!

need information on shipping specifics for components and storage temp requirements?
The shipping specifics and storage temperature requirements for the components are as follows: - Component 632637, which is part of the product 632635, should be stored at -20°C and shipped with dry ice. - Component 632638, also part of product 632635, can be stored at room temperature and should be shipped at room temperature. In summary, product 632635 consists of both components: 632637 (shipped cold) and 632638 (shipped at room temp).

hey, how can i find out the expiration or lot info for my rri labels since we tossed the packaging?
To find out the expiration or lot information for your RNase Inhibitor labels after disposing of the packaging, you can reach out to the manufacturer or supplier directly. They typically keep records associated with each lot number and can provide you with details like the expiration date. If possible, include any relevant information such as the product name and lot number in your inquiry to expedite their response.

Could you please clarify the proper format for my email address?
It seems that there was an issue with the email you attempted to send, and it bounced back. If you're looking for clarification on the correct format for your email address, ensure that it includes the appropriate components: a username followed by "@" and then the domain name (e.g., example.com). Double-check for any typos or missing elements. If needed, feel free to reach out again for assistance!

shelf life of lenti-x lenti concentrator?
The shelf life of the Lenti-X Lenti Concentrator is lot-specific, with an internal shelf life of 3 years from the manufacturing date. If you need further assistance or have more questions, feel free to ask!

why didn't I receive the original email to create my research account?
There could be several reasons why you didn't receive the original email to create your research account. It's possible that it may have ended up in your spam or junk folder, so checking there is a good first step. If you still can't find it after checking, consider reaching out to the support team that handles research and purchasing accounts for further assistance. They should be able to help you with any issues related to receiving important emails or initiating your account application process.

how to ensure optimal rna yield without damaging fragile cells during virus addition?
To ensure optimal RNA yield while protecting fragile cells during virus addition, consider the following steps: 1. **Coating Protocol**: Use RetroNectin at the recommended concentration (4-20 µg/cm²) to coat your plates, ensuring sufficient binding for effective transduction. Following this, counter-coat with BSA to minimize uncoated areas that could interfere with the process. 2. **Centrifugation Conditions**: If centrifugation is necessary to facilitate virus binding, adjust parameters based on cell fragility. For sensitive cells, you may want to reduce g-force and duration or omit centrifugation altogether if possible. 3. **Volume Management**: Use appropriate volumes of virus solution—generally no more than 2 ml for a 6-well plate—to prevent drying out during incubation and ensure even distribution over the surface area. 4. **Maintain Sterility**: Covering plates with cling film or parafilm during any centrifugation step can help maintain sterility and avoid aerosolization of viral particles. 5. **Gentle Handling Techniques**: When adding viruses or cells post-centrifugation, use gentle pipetting techniques to minimize shear stress on fragile cells. By carefully balancing these factors, you can enhance RNA yield while minimizing damage to delicate cell types during viral transduction processes.

can you clarify the role of RACE CDS primers in the cDNA synthesis and how they differ from oligo dT primers?
RACE CDS primers play a specific role in the synthesis of cDNA by facilitating the amplification of RNA sequences, particularly when trying to obtain complete transcripts. The 5' RACE CDS Primer A is an Oligo(dT) that helps generate first-strand cDNA from mRNA with a polyA tail but does not contain any additional primer binding sites. This allows it to anneal specifically to the polyA tail. In contrast, the SMARTer II A oligo incorporates a primer binding site at the 5' end of full-length cDNA through template switching activity during reverse transcription. This feature enhances subsequent amplification and cloning processes. On the other hand, the 3' RACE CDS Primer A is also an Oligo(dT) but includes specific primer binding sites designed explicitly for use in 3' RACE applications. Its primary function is to allow for targeted amplification of RNA sequences starting from their polyA tails while ensuring compatibility with downstream processes. Overall, while both types of primers are involved in cDNA synthesis related to mRNA transcripts, their structures and functions differ significantly based on whether they are used for general first-strand synthesis or specialized applications like RACE.

retronectin photo sensitive?
The conversation indicates that T202, which is kept in clear glass vials, does not exhibit photosensitivity. If it were photosensitive, it would typically be packaged in a colored vial to protect it from light exposure. However, there are concerns regarding the quality of the product when stored at room temperature for longer than 12 hours; while the product may still be functional, its GMP quality cannot be assured under those conditions.

what is the estimated total time length for library prep using a total rnaseq kit with ribodepletion step and rid globin mrna side?
The estimated total time length for library preparation using a total RNA-seq kit with a ribodepletion step, as described in the provided overview of the SMART-Seq Total RNA Pico Input with UMIs (ZapR Mammalian) protocol, is approximately 6 hours and 30 minutes. However, actual processing time may vary based on factors such as the number of samples being processed and specific cycling conditions used during PCR. Additionally, it is noted that this particular kit does not remove globin RNA from the sample.

happen know ready-to-use split fluorescent proteins for research?
There are indeed ready-to-use split fluorescent proteins available for research purposes. These typically come in two halves, which can be used to study protein interactions and localization by reconstituting fluorescence when the two halves come together within a biological system. If you're looking for specific products or suppliers, it might be helpful to check with specialized biotechnology companies that offer these tools. Additionally, some resources may provide protocols for generating your own split versions from existing fluorescent proteins like GFP if you prefer a custom approach.

do i need to have two thermal cyclers for the smart-seq protocol or can one thermocycler work adequately
It is generally recommended to use two thermal cyclers for the Smart-Seq protocol to prevent contamination; however, if you only have one available, it can still work adequately with careful precautions. Ensure that you thoroughly clean the thermal cycler before starting cDNA synthesis or PCR amplification. Additionally, placing the thermal cycler in a clean room can help minimize contamination risks and improve results.

how does the probe design ensure that only mammalian rRNA is depleted while leaving other transcripts intact?
The probe design specifically targets sequences that are unique to mammalian rRNA, allowing for effective depletion of these transcripts while minimizing the impact on other RNA species. Since the probes are designed to bind selectively to mammalian rRNA and not to other types of RNA, this specificity helps ensure that non-mammalian or different transcript categories remain intact. The use of N6 random primers in reverse transcription further supports this by providing a broad representation without bias toward any particular species, which is advantageous for obtaining a more comprehensive view of the remaining transcriptome after depletion.

how to check illumina license status?
To check the license status for Illumina, it is recommended to reach out directly to customer service. They can provide specific information regarding your order and any licensing requirements. Additionally, you may contact the licensing department of the relevant company for further clarification on licenses associated with their technologies. If you're affiliated with a for-profit organization or have questions about commercial use, it's essential to inquire about obtaining a commercial license agreement as well.

where can i find data sheets for the origin of total rna?
To find data sheets for the origin of total RNA, you can visit the manufacturer's website or specific product pages where they typically provide certificates of analysis and related documentation. If you're looking for certain lot numbers or products, searching through their "Documents" tab may be beneficial. For any specific inquiries about particular products, it might also be helpful to contact customer support directly on the website for more tailored assistance.

do you offer support with writing?
Yes, support with writing is available. If you have specific needs or projects in mind, feel free to share the details, and assistance can be provided to help you achieve your goals.

Would using SPRI beads instead of NucleoMag beads in the Takara Total RNA High Input RiboGone library prep kit affect my library yield compared to the recommended Ampure beads in the Smart-Seq Stranded Total RNA-seq Pico Input Kit?
Using SPRI beads instead of NucleoMag beads in the Takara Total RNA High Input RiboGone library prep kit may negatively impact your library yield. Customer feedback has indicated that SPRI beads can lead to lower yields for various libraries. In contrast, AMPure XP PCR purification kits are recommended and considered appropriate substitutes if NucleoMag products are unavailable. Therefore, it would be advisable to stick with AMPure beads for optimal results in your library preparation.

What are the return policies for items bought online?
Return policies for items bought online typically vary by retailer, but common practices include allowing returns within a specified period—often 30 days from the date of purchase. Items usually need to be in their original condition and packaging to qualify for a refund or exchange. Some retailers may provide free return shipping, while others might require customers to cover the cost. It's important to check the specific return policy on the retailer's website or contact customer service for detailed information regarding your order.

What are your hours of operation?
The hours of operation are not specified in the provided conversation. It would be best to check directly with the relevant organization or their website for accurate and up-to-date information regarding their hours.

What specific type of FBS do you recommend for the protocol?
For the protocol, it is recommended to use FBS that is specifically labeled as "Tet-approved." This type of FBS is designed to minimize any potential complications from tetracycline antibiotics, which can interfere with your packaging system that utilizes Tet-Off technology. Using this approved serum will help ensure optimal cell performance and higher titers in your experiments.

umi's kit smart-seq smart seq mrna lp umis resulting sequencing analyzed without cogent pipeline open source?
The SMART-Seq Pico kits utilize unique molecular identifiers (UMIs) that enhance the accuracy of RNA sequencing by allowing for the identification and removal of duplicate reads. While Cogent is recommended for its effective mapping and UMI collapsing capabilities, you can still analyze sequencing data generated from SMART-Seq without using the Cogent pipeline. Open-source alternatives are available that can handle UMI processing, such as tools like UMI-tools or other RNA-seq analysis pipelines that support UMI-based methods. However, keep in mind that these may require additional steps to ensure proper handling of duplicates compared to what Cogent offers out-of-the-box. It's essential to review your specific analysis goals and consider how you will manage UMIs during preprocessing if not utilizing the Cogent pipeline. Properly implementing an alternative method will help maintain data integrity while analyzing your results effectively.

Are the primers I designed using Primer Blast compatible with the Takara in-fusion cloning kit?
The primers you designed using Primer Blast can be compatible with the Takara In-Fusion Cloning Kit, provided they follow the specific guidelines outlined in the In-Fusion HD Cloning Kit User Manual. It's essential to ensure that your primers meet the design parameters mentioned in section IV B of the manual, such as length, melting temperature (Tm), and GC content. After designing your primers manually or with a tool, it's important to perform a BLAST search to confirm that your primers do not anneal to similar sequences on your template. This step is crucial for ensuring specificity during PCR amplification. Once you've confirmed these aspects, you should be set for use with the In-Fusion cloning process.

best kit rna isolation ivt?
For RNA isolation in an in vitro transcription (IVT) context, the choice between the LiCl precipitation method and the NucleoSpin® RNA Clean-up Maxi kit depends on your specific requirements. If you are dealing with larger RNA sizes (greater than 300 bases) and concentrations above 0.1 μg/μl, LiCl precipitation can effectively remove uncaptured NTPs and proteins. However, if your RNA size is smaller or concentration is below 0.1 μg/μl, using the NucleoSpin® RNA Clean-up Maxi kit would be more suitable as it allows for efficient purification even under these conditions. Ultimately, both methods have their advantages depending on the characteristics of your sample. If you're looking for a reliable option that covers a wider range of scenarios including lower concentrations and smaller fragments, then the NucleoSpin® RNA Clean-up Maxi kit would be recommended for IVT applications.

wonder why bands equal intensity mean positive negative strains?
The intensity of bands on a Western blot can indicate the relative abundance of the protein being analyzed. If you observe equal intensity bands in both positive and negative control strains, it could suggest that there is background noise or non-specific binding occurring. It may also imply that the untransformed cells are expressing some level of endogenous protein that cross-reacts with your antibody. To better understand this phenomenon, consider optimizing your experimental conditions, such as antibody dilution or blocking steps, to reduce background signal and ensure specific detection. Additionally, comparing samples from different cell lines or using alternative antibodies might help clarify the results.

hi, can you provide the contact email for the e-procurement procurement team?
You can reach out to the e-procurement team by contacting their designated email address. If you need further assistance or have any other questions, feel free to ask.

why is there no visible pellet after using lenti-x concentrator with serum free medium?
The absence of a visible pellet after using the lenti-x concentrator with serum-free medium can be attributed to several factors. First, the pellet may not always appear white or opaque; it can often be translucent due to the nature of the components in the serum or medium used. Additionally, if there was residual medium left behind during processing, it could dilute any virus that might have precipitated, leading to little visibility of a pellet. It's also important to ensure that sufficient dissolving buffer is added after centrifugation. This helps bring any potentially invisible virus into solution from the pellet on the tube wall. The efficiency of resuspension can be affected by how much residue remains in the tube; therefore, removing as much as possible without losing viral content is crucial. Lastly, even if there isn't an apparent visible pellet, this does not necessarily indicate an absence of virus—it's possible for viruses to still be present in small amounts within what appears as a translucent substance or attached to surfaces within the tube.

what should i do if my cloning resulted in the backbone vector being changed and important parts missing?
If your cloning resulted in the backbone vector being altered and important sequences like the T7 promoter and BGHR missing, consider taking the following steps: 1. **Verify Results**: Confirm that the deletions are consistent across multiple colonies. Sequence additional clones to assess whether this was an isolated incident or a common issue. 2. **Review Cloning Protocol**: Double-check your PCR amplification process for both the insert and linearized vector. Ensure that primers were designed correctly with appropriate overhangs for In-Fusion cloning. 3. **Linearization Method**: Evaluate how you linearized your vector—whether through PCR or restriction digestion—and ensure that this method did not introduce unintended deletions. 4. **Check Competent Cells**: If you transformed into Stellar competent cells, verify their efficiency and consider using another strain if necessary, as some strains may have issues with specific plasmids. 5. **Reduce Growth Conditions**: As high-copy vectors can be sensitive during replication, try culturing at a lower temperature (30-34°C) to reduce stress on the plasmid which could lead to instability. 6. **Consider Other Factors**: Investigate if there are any repeat sequences present in your vector that might contribute to recombination events leading to deletions during culture growth. 7. **Replicate Successful Clones**: If other clones showed no deletions, prioritize those for further experiments while troubleshooting this particular clone's issues. 8. **Documentation of Findings**: Keep detailed records of all attempts, findings, and methods used so you can analyze patterns or share information effectively if seeking help from others in your lab or community. By systematically addressing these areas, you should gain insight into what went wrong and how to prevent similar issues in future cloning efforts.

proper recommendation wanted confirm?
For exosomal RNA profiling, it is indeed recommended to use SMART-Seq Pico kits due to their effectiveness in preparing high-quality libraries from small amounts of RNA. It's crucial that all traces of DNA are thoroughly removed during the purification process, as residual single-stranded DNA can co-purify with RNA and interfere with downstream applications. In studies such as those by Yonghui Su et al. and Anna Almeida et al., DNase treatment was employed to eliminate any contaminating DNA before constructing strand-specific RNA-seq libraries. This step is essential for ensuring accuracy in the analysis of long RNAs derived from extracellular vesicles (EVs), particularly when assessing biomarkers related to conditions like breast cancer or prostate cancer. Additionally, confirming the DV200 metric of purified RNA will help ensure its integrity for sequencing purposes. Overall, following these guidelines will enhance the reliability and relevance of findings in exosomal RNA research.

could switching to serum-free medium affect the visibility of the pellet after using lenti-x lentiviral sup?
Switching to serum-free medium can indeed affect the visibility of the pellet after using lenti-x lentiviral supernatant. In serum-containing media, various proteins and components contribute to the appearance of the pellet, often making it more opaque or white. However, in a serum-free medium, you may notice that the pellet could appear more translucent or even different in color due to the absence of those additional proteins. This change does not necessarily indicate any loss of viral particles; rather, it reflects how different mediums impact protein precipitation and pellet characteristics. Therefore, while visibility might be affected, it's essential to consider that a translucent pellet doesn't imply a lack of virus present.

is rectronectin coating ideal for transducing neurons, astrocytes, and microglia in matrigel-based ECM?
Using RetroNectin coating for transducing neurons, astrocytes, and microglia in a matrigel-based ECM may not be ideal. Matrigel can interfere with the binding of cells and viruses to RetroNectin, which is crucial for enhancing transduction efficiency. Additionally, it's important to consider that coated plates generally reduce the effectiveness of RetroNectin compared to non-treated plates. For optimal results, it is recommended to use non-coated tissue culture plates or dishes when utilizing RetroNectin for cell transduction.

do rectronectin coatings affect the survival of transduced neurons, astrocytes, and microglia in matrigel based ecms?
RetroNectin coatings can impact the transduction efficiency of neurons, astrocytes, and microglia in Matrigel-based ECMs. Since RetroNectin facilitates binding between cells and the virus for enhanced transduction, its effectiveness may be compromised by factors present in Matrigel. It's important to consider that while RetroNectin enhances transduction efficiency, the presence of Matrigel could potentially affect cell yield and viability. Conducting comparative studies with different conditions could help determine how these factors interact and influence the survival of your specific cell types post-transduction.

what is the difference in rnase activity between smartscribe reverse transcriptase and smart mmlv reverse transcriptase, and how does it affect template switching and cdna generation?
The main difference in RNase activity between SmartScribe Reverse Transcriptase and SMART MMLV Reverse Transcriptase lies in the level of RNase H activity. SmartScribe RT has reduced RNase H activity, which is beneficial for template switching applications as it minimizes degradation of the RNA template during reverse transcription. This allows for more efficient incorporation of additional sequences onto the 3' end of cDNA. In contrast, SMART MMLV RT retains higher levels of RNase H activity, which can be advantageous for generating high-quality cDNA from RNA templates but may not be as effective for template switching due to potential degradation of the RNA strand. While both enzymes are capable of producing first-strand cDNA, those requiring efficient template switching often prefer SmartScribe due to its reduced RNase H effect. Additionally, each enzyme supports different lengths of cDNA synthesis: SMART MMLV RT can produce up to 11 kb products while SmartScribe RT can generate longer fragments up to 14 kb.

sybr green what is it and how do I use it?
SYBR Green is a fluorescent dye commonly used in quantitative real-time PCR (qPCR) to detect and quantify DNA. It binds to double-stranded DNA, emitting fluorescence when excited by light, allowing for the measurement of DNA amplification during PCR cycles. To use SYBR Green effectively: 1. **Prepare Your Reaction Mix**: Include your target primers, template DNA, dNTPs, buffer solution, and the SYBR Green reagent in your qPCR reaction mixture. 2. **Set Up PCR Conditions**: Follow a thermal cycling protocol that typically includes denaturation (usually around 95°C), annealing (specific temperature depending on primer design), and extension (generally at 72°C). 3. **Use Appropriate Controls**: Include no-template controls to check for contamination and positive controls to validate your assay's efficiency. 4. **Analyze Results**: After completing the PCR cycles, analyze the data using a real-time PCR machine capable of detecting SYBR Green fluorescence at each cycle. This method can be combined with various qPCR reagents like TB Green® from Takara or others specified for intercalator-based detection systems. Always refer to specific product manuals for detailed instructions tailored to those products.

think enough treatment clean dna again prior infusion step or should i do something else?
It's important to ensure that your DNA is clean before proceeding with the In-Fusion step. If you've confirmed a single band on the gel, you can either remove the template DNA using Cloning Enhancer or perform additional cleanup if needed. Since DpnI isn't used in In-Fusion reactions and may affect them due to its buffer components, sticking with Cloning Enhancer would be a suitable option. If you're uncertain about any residual contaminants, it might be beneficial to clean up the sample again for optimal results prior to infusion.

what are the details of alternate verifiler express pcr reagents?
Currently, there are no direct alternatives to the VeriFiler Express PCR Amplification Kit that provide the same capabilities for direct amplification of DNA from common sample types without extraction or purification. The VeriFiler Express kit is specifically designed for amplifying single-source human genomic DNA and offers a 25-locus multiplex with high discrimination power, making it ideal for paternity testing and similar applications. While there may be other forensic DNA reagents available in the market, they might not have been validated for use with specific sample types like buccal samples on treated or untreated paper or blood samples. If you're considering different options such as Terra Direct, it's important to note that validation with short tandem repeat (STR) analysis has not been established yet. Overall, if you require an alternative product, it would be advisable to consult directly with manufacturers or suppliers who specialize in forensic DNA analysis kits to explore any new developments or offerings.

how does the bcr enhancer work to improve low RIN RNA samples?
The BCR enhancer is designed to enhance the RT (reverse transcription) reaction, which can be particularly beneficial when working with RNA samples that have low integrity. However, it's important to note that specific details about how the BCR enhancer operates are proprietary and not publicly available. For optimal results, using the BCR profiling kit requires high-quality RNA with a RIN (RNA Integrity Number) greater than 7. Low RIN RNA is incompatible because: - 3'-end truncated RNA may not be properly dT primed during reverse transcription, leading to reduced representation in the final BCR library. - 5'-end truncated RNA might miss critical V(D)J sequences located at the mRNA's 5' end, which can hinder accurate clonotype identification. Thus, while the BCR enhancer aims to improve reactions for low quality samples generally, it ultimately works best when starting with high-integrity RNA.

what are the best strategies to improve efficiency of sgRNA transduction in iPSCs and their differentiated cells?
To improve the efficiency of sgRNA transduction in iPSCs and their differentiated cells, consider the following strategies: 1. **Use of Lenti-viral Vectors**: Lentiviral vectors are effective for gene delivery as they integrate into the host genome and can target actively transcribed genes. Their ability to provide stable expression makes them suitable for both iPSCs and their differentiated derivatives. 2. **Optimizing Coating Techniques**: When using retronectin, ensure proper coating of plates at 4-20 µg/cm² to enhance viral binding. For example, preparing a 20 µg/ml solution and applying it correctly will help achieve an adequate density on your culture surface. 3. **Infection Methods**: Consider different infection methods such as the RetroNectin-Bound Virus (RBV) method where you coat plates with retronectin before adding virus or mixing virus with cells prior to plating them onto coated wells. Choose based on which gives better results in your specific context. 4. **Transduction Conditions**: Optimize conditions like multiplicity of infection (MOI), incubation time, and temperature during transduction to maximize uptake by target cells. 5. **Post-transduction Selection**: Implement a selection process post-transduction if applicable, using antibiotic resistance markers or fluorescence-based sorting techniques that allow enriched populations expressing your sgRNAs effectively. 6. **Monitoring Integration Efficiency**: Analyze integration sites and copy number after differentiation since these parameters may influence long-term expression levels of sgRNA constructs. 7. **Utilizing Different Viral Systems**: If lentivirus does not yield desired efficiency, explore other viral systems like AAVs or adenoviruses that might offer higher transduction rates depending on cell type specificities. These approaches combined can significantly enhance your sgRNA transduction efficiency in both undifferentiated iPSCs and their differentiated forms while maintaining good experimental reproducibility.

is there a specific reason why step PCR is more effective for annealing than regular cycling?
Step PCR, which typically involves a three-step cycling process (denaturation, annealing, and extension), is often more effective for annealing compared to regular cycling because it allows for precise control over the temperature and duration of each step. The separate annealing phase provides adequate time for primers to bind specifically to their complementary sequences on the template DNA. This increased specificity can be particularly beneficial when dealing with complex templates or GC-rich regions where non-specific binding might occur. In contrast, regular cycling may have shorter or less distinct phases that could lead to inefficient primer binding or increased non-specific amplification. By optimizing each step independently, including using an appropriate temperature gradient during the annealing phase based on the melting temperatures (Tm) of the primers, step PCR enhances specificity and overall yield in amplifying target sequences.

can anyone help me with the math for these ratios and products in my sales agreement?
To assist with the calculations for your sales agreement based on the provided information, let's break down some key components and their quantities: 1. **Recombinant RNase Inhibitor (40 U/µl)**: For a 24-reaction format of the SMART-Seq® Human BCR (with UMIs), you noted that it includes 24 µl of RNase Inhibitor. If you need more than this amount, consider purchasing additional units like product 2313A, which comes in 5,000 U at a concentration of 40 U/µl. This yields: - Total volume needed = \( \frac{5000 \text{ U}}{40 \text{ U/µl}} = 125 \text{ µl} \) 2. **SMARTScribe Reverse Transcriptase**: The protocol indicates you'll require a total of 48 µl for your reactions using SMARTScribe RT at a concentration of 100 U/µl. You can purchase product number 639537 which provides you with a total volume of: - Volume purchased = \(100\) µl 3. **PrimeSTAR GXL Premix (2X)**: Each PCR reaction requires 25 µl per reaction for both PCR1 and PCR2 stages; therefore, for two rounds in one set: - Total volume required = \(25\) µl * \(2\) reactions * \(24\) samples = \(1200\) µl To fulfill this requirement efficiently, consider purchasing product R051A as it offers multiple sizes. If you're looking to optimize your reagent quantities or have specific ratios in mind related to sample numbers or other conditions not mentioned here, sharing those details will enable more precise calculations tailored to your project needs.

What can I do to make this day even greater?
To make your day even greater, consider engaging in activities that bring you joy and fulfillment. You could take a moment to reflect on what you're grateful for, spend time outdoors enjoying nature, or connect with friends or loved ones. Engaging in a hobby you love, practicing mindfulness or meditation, or treating yourself to something special can also enhance your day. Prioritizing self-care and positivity will contribute significantly to making today memorable.

do i need extra gel trays and combs for the mupid-exu set?
The AD140 Mupid exU typically includes a complete set of components, but whether you need extra gel trays and combs depends on your specific usage needs. If you anticipate frequent use or require backups for different experiments, it might be beneficial to have additional gel trays and combs on hand. Checking the user manual you received could also provide further clarification on this matter.

how can i improve the efficiency of re-transducing sgrna in ips cells for better differentiation outcomes?
To improve the efficiency of re-transducing sgRNA in iPS cells for better differentiation outcomes, consider optimizing several key factors. First, assess the timing and conditions of transduction to ensure that it coincides with the right stage of cell cycle when your iPS cells are most amenable to integration. Additionally, you might explore varying the multiplicity of infection (MOI) to find an optimal level that maximizes uptake without overwhelming the cells. Utilizing lentiviral vectors can also be beneficial since they integrate into actively transcribed regions, which may enhance expression stability. You could experiment with different promoters or enhancers in your constructs that drive expression more effectively during differentiation phases. Furthermore, incorporating small molecules or signaling compounds known to support pluripotency maintenance and transition towards specific lineages can help create a more favorable environment for successful differentiation post-transduction. Lastly, monitoring transgene expression levels after integration will provide insights into potential adjustments needed in protocol parameters for improved outcomes.

is smartseq mrna lp kit suitable for low concentration rna with poor rin scores?
The SMART-Seq mRNA LP kit, which requires high integrity RNA with a RIN score greater than 8, may not be suitable for low concentration RNA with poor RIN scores. For such samples, the SMART-Seq® Total RNA Pico Input with UMIs (ZapR® Mammalian) could be a better option as it utilizes random priming for library construction and is designed to accommodate lower quality RNA. However, it's important to ensure that the DV200 metric is determined and that all traces of DNA are removed before proceeding.

Can you provide more details about the process?
The NucleoBond® Xtra Maxi Columns are designed for efficient nucleic acid purification. The process typically involves the following steps: 1. **Cell Lysis**: Cells containing the nucleic acids are lysed to release DNA or RNA. 2. **Binding**: The lysate is applied to the column, where nucleic acids bind specifically to the resin in the column while impurities pass through. 3. **Washing**: After binding, a wash buffer is used to remove contaminants and proteins from the bound nucleic acids. 4. **Elution**: Finally, an elution buffer is added to release purified nucleic acids from the column. These columns are available individually or in bulk quantities (10 or 100 columns), which allows flexibility depending on your experimental needs. For more detailed instructions tailored to specific applications, it's recommended to refer directly to product documentation provided by Takara Bio on their website.

so is there a way to get the full experiment done without purchasing more tet fbs?
The initial 50 ml aliquot provided with the tet systems is typically just enough to prepare one bottle of media, meaning that additional tet FBS would be necessary to complete a full experiment. While other types of FBS may work, our specific formulation has been tested to ensure compatibility with the tet system, so it’s recommended to use it for reliable results.

hello which polymerase has the highest fidelity for long sequencing?
The polymerase with the highest fidelity for long sequencing among those listed is the PrimeSTAR Max DNA Polymerase, which has a fidelity of 0.0002%, equating to 465 times that of Taq polymerase.

what can i do to improve the quality of my sequencing data and avoid unannotated reads?
To improve the quality of your sequencing data and minimize unannotated reads, consider the following steps: 1. **Use Appropriate Libraries**: Ensure you are using the SMART-Seq® Total RNA Pico Input with UMIs (ZapR™ Mammalian) correctly, as this method incorporates unique molecular identifiers (UMIs) to help identify and quantify transcripts more accurately. 2. **Spike-in Control**: It is recommended to include 10% PhiX spike-in when running the NextSeq 2000. This can enhance clustering during sequencing and improve overall data quality. 3. **Check Trimming Settings**: Verify whether automatic adapter trimming is enabled on your sequencer settings. If not, ensure that any necessary adjustments are made to avoid issues related to adapters interfering with read quality. 4. **Utilize Cogent NGS Analysis Pipeline**: Employ this free-of-charge tool from Takara Bio for processing your raw non-demultiplexed sequencing reads, as it collapses UMIs and maps them effectively. 5. **Validate RNA Samples**: Prior to library construction, validate that your RNA samples contain an appropriate small RNA fraction if that's relevant for your specific analysis goals. 6. **Consider Size Selection**: Implement size selection carefully during library preparation; this helps in enriching for desired transcript sizes while eliminating unwanted products that may contribute to unannotated reads. 7. **Understand Library Composition**: Be aware that some small RNAs may originate from introns of coding mRNAs or other sources within total RNA input types—this knowledge can guide further analyses or interpretations of results. 8. **Monitor UMI Linker Effects**: To alleviate low diversity introduced by UMIs, make sure there’s a proper design involving nontemplated nucleotides derived from the UMI linker in order to maintain sufficient complexity in libraries generated. By taking these measures into account throughout sample preparation and data analysis processes, you should see improvements in both sequencing quality and annotation accuracy of your resulting datasets.

how does the performance of the dna smart chip-seq kit compare to the thruplex kit in terms of data sensitivity?
The DNA SMART ChIP-Seq Kit and the ThruPLEX kit serve different purposes and have unique characteristics that affect their performance, particularly in terms of data sensitivity. The DNA SMART ChIP-Seq Kit is designed specifically for chromatin immunoprecipitation (ChIP) applications. It allows for the sequencing of both single-stranded (ssDNA) and double-stranded (dsDNA) inputs, which can enhance its sensitivity by enabling it to capture a broader range of epigenetic modifications. On the other hand, the ThruPLEX kit is optimized for library preparation from dsDNA only. This limitation may impact its overall sensitivity when working with samples where ssDNA might provide additional insights or better recovery rates. In conclusion, while both kits can produce high-quality sequencing results, the DNA SMART ChIP-Seq Kit has an advantage in terms of flexibility with ssDNA input, potentially leading to enhanced data sensitivity in specific applications related to ChIP-seq workflows compared to the ThruPLEX kit.

Greetings
Hi $$$visitor_first_name$$$. Thank you for reaching out! Please let me know how I may help you today.

email to reach customer
Is this the best email to reach you? $$$Company_Email$$$

Greetings
Hi $$$Operator_First_Name$$$. Thank you for reaching out! Please let me know how I may help you today.

what should I do on my rest day?
On your rest day, consider engaging in activities that promote relaxation and recovery. You could take a leisurely walk or go for a gentle bike ride to enjoy some fresh air. Practicing mindfulness through meditation or yoga can also help rejuvenate your mind and body. If you prefer something more enjoyable, reading a book, watching a movie, or trying out new recipes might be great options. Additionally, spending time with friends or family can provide social fulfillment while allowing you to unwind. Prioritize whatever helps you feel restored and ready for the upcoming days.

freeze thaw cycles lentivirus safe long keep packaged activity
Freeze-thaw cycles can significantly affect lentivirus titers, typically reducing them by about 50% with each cycle. To maintain the activity of lentiviruses during storage, it's advisable to aliquot your samples and avoid repeated freeze-thawing. When using a concentrator, it is recommended to store the virus at 4°C for no more than 12 hours without it; otherwise, prolonged periods should be avoided due to the fragility of lentiviruses. Additionally, ensuring that packaging cells are healthy—ideally between passages 5 and 25 after thawing—and using high-quality endotoxin-free viral vector DNA are key factors in preserving viral activity over time.

how does takara's smrna-seq kit compare to diagenode's d-plex small rna-seq kit and illumina's rip-seq kit in detecting low abundance small rnas?
Takara's SMARTer smRNA-Seq Kit is specifically designed for small RNA profiling, allowing for input amounts ranging from 1 ng to 2 μg. This kit is particularly effective in detecting low-abundance small RNAs due to its sensitivity and optimization for this purpose. In comparison, Diagenode's D-Plex Small RNA-Seq Kit also targets low abundance small RNAs but may have different workflow specifications or performance metrics depending on sample type and quality. The D-Plex kit is known for enabling multiplexing of samples, which can enhance throughput while maintaining sensitivity. Illumina's RIP-seq kits are tailored more towards the analysis of RNA-protein interactions rather than solely focusing on small RNAs. While they can effectively detect small RNAs within the context of m6A methylation studies (as shown in the references you provided), their primary design caters to capturing immunoprecipitated RNA fragments, which might not be as optimized purely for small RNA detection compared to dedicated kits like those from Takara or Diagenode. Overall, if the goal is purely focused on detecting low-abundance small RNAs with a streamlined approach, Takara’s SMARTer smRNA-Seq Kit may provide a more targeted solution compared to Illumina’s RIP-seq offerings; however, comparing it directly with Diagenode would require consideration of specific experimental needs and parameters such as yield requirements and library preparation protocols.

what can i do to make sure my pcr premix stays viable after the delivery issues?
To ensure your PCR premix remains viable after experiencing delivery issues, consider the following steps: 1. **Storage Conditions**: Immediately check the storage temperature upon receipt. Ensure that the premix is stored at the recommended temperature, typically -20°C or -80°C for long-term storage. 2. **Control PCR**: Run a control PCR to test the functionality of your polymerase and other components in the mix. This will help confirm whether they are still active despite any potential thermal exposure during transit. 3. **Avoid Freeze-Thaw Cycles**: If possible, minimize freeze-thaw cycles by aliquoting your premix into smaller portions before freezing, so you only thaw what you need for each experiment. 4. **Check Expiration Dates**: Verify that all reagents are within their expiration dates to ensure optimal performance. 5. **Monitor Temperature During Use**: When preparing reactions, keep reagents on ice until ready to use to maintain stability. By taking these precautions, you can enhance the viability of your PCR premix and optimize results in your experiments.

is there an issue with the phone lines?
There doesn't appear to be any issues with the phone lines for customer service, as they are open for calls. If you've had trouble reaching them, please feel free to provide your contact number so that someone can follow up with you directly.

why vk constant region contains stop code in sequencing results?
The presence of stop codons in the constant region sequences from your sequencing results can be attributed to the library construction process used in the SMARTer® Mouse BCR IgG H/K/L Profiling Kit. During this process, additional sequences are introduced that may inadvertently create stop codons. These added sequences are not relevant for sequencing the V(D)J region and can lead to artifacts when interpreting results. It's important to note that while we analyze primarily the V(D)J regions for clonotype identification, we do not validate or focus on the constant region during our assessments. The extra sequences at both ends of mRNA and next to the constant region could contribute to these unintended stop codons observed in your data.

rna immunoprecipitation rip low input kit recommend?
For RNA immunoprecipitation (RIP) with low input, the SMARTer Stranded Total RNA-Seq Kit v2-Pico Input Mammalian is highly recommended. This kit is designed for low-input samples and allows for effective construction of libraries from small amounts of RNA, such as 10 ng fragmented RNA. It has been successfully utilized in studies involving m6A MeRIP-seq and other epitranscriptome analyses. Additionally, considering methods that refine RIP protocols for low input materials can enhance your results. Literature references such as those by Xia et al., which detail using the SMARTer Stranded Total RNA-Seq Kit alongside specific immunoprecipitation techniques, provide valuable insights into successful applications. If you have any further questions or need more information on this topic, feel free to ask!

what is the estimated concentration of guanidinium thiocyanate in lysis buffer and how does it compare to SDS?
The estimated concentration of guanidinium thiocyanate in the Lysis Buffer (LBP) is about 45%. This concentration is significant as it serves as a strong denaturant, effectively inactivating viruses by lysing envelope proteins and other viral components. In comparison to SDS (sodium dodecyl sulfate), which is commonly used for protein denaturation, guanidinium thiocyanate provides a different mode of action due to its ability to disrupt hydrogen bonds and solubilize proteins. While both agents are effective in their roles, they operate through distinct mechanisms that can be advantageous depending on the specific application or protocol being utilized.

what's the typical time for a customer to do single wash strips cut etc?
The typical time for a customer to prepare single wash strips, including cutting and positioning them for the plate reader, can vary based on the individual's familiarity with the process and setup. However, generally speaking, customers should allocate some extra time for these tasks to ensure proper alignment and effective washing steps. It’s important that they feel comfortable with each step to achieve optimal results. If they're working with multiple samples or need to create standard curves, managing their workflow efficiently will also help in reducing overall processing time.

when will the customer receive protocol access?
The customer will receive protocol access once the necessary checks have been completed and the source has been consulted. An email update will be sent later to provide further details on the timeline.

am hoping to know what the expected recovery percentage of viral RNA is when using the nucleospin RNA virus kit from Takara?
The expected recovery percentage of viral RNA when using the NucleoSpin® RNA Virus kit from Takara can vary based on several factors, including the initial viral titer and the type of sample being processed. Generally, specific recovery rates are not provided in product documentation as they can be influenced by sample composition and handling methods. For precise measurements, it's recommended to conduct an RT-qPCR assay to quantify the amount of RNA recovered after purification. If you have a known concentration or titer of your starting material, it would help in estimating recovery more accurately.

Can you send me the documents for the lentivirus titration kit by email?
The documents for the Lenti-X qRT-PCR Titration Kit have been sent via email. If you need anything else or have further questions, feel free to ask!

know any similar site available us
There are several sites available in the U.S. that offer similar genomic DNA purification kits. Some reputable options include: 1. **Thermo Fisher Scientific** - They provide a variety of DNA extraction and purification products suitable for different sample types. 2. **Qiagen** - Known for their comprehensive range of molecular biology kits, including those specifically designed for tissue samples. 3. **Promega** - Offers various nucleic acid purification solutions that may meet your needs. 4. **Bio-Rad** - Provides kits focused on nucleic acid isolation from various biological samples. These companies have extensive product catalogs online where you can find detailed information about their offerings to see which best fits your requirements.

order old kit for bacteria colonies?
The In-Fusion® HD Cloning Kit (639649) may still be available for order. However, it's worth noting that the HD kit has been replaced by the In-Fusion Snap Assembly Master Mix, which improves colony generation. To check on the availability of the old kit and place an order, please reach out to customer service directly via email or phone. They will provide you with the necessary assistance regarding your order request. If you have any other questions or need further help, feel free to ask!

what happens to gmp grade retronectin if stored at warmer temperatures?
Storing GMP grade retronectin at warmer temperatures than the recommended storage temperature of -30°C can lead to a loss of activity and potentially invalidate assay processes. Since retronectin is a protein sensitive to temperature changes, deviations from proper storage conditions may compromise its effectiveness. It is crucial to maintain the specified storage conditions to ensure that the product remains viable for use in your applications. If there are any concerns about handling or shipping, it’s advisable to communicate with the supplier for further guidance on ensuring optimal conditions during transport and usage.

what is the recovery value of viral rna isolated using nucleospin rna virus kits?
The recovery value of viral RNA isolated using the NucleoSpin RNA Virus kits can vary, and it's often challenging to provide a specific estimate without knowing the initial viral titer or copies. These kits are primarily designed for isolating viral RNA from biological fluids like serum, urine, and swab washes, but they do not quantify recovery directly. To assess the actual amount of recovered viral RNA, you would typically use RT-qPCR methods. However, it’s essential to note that while these assays can measure the presence of RNA, not all isolated RNA will be packaged into infectious viruses. Therefore, quantifying physical titer may require additional steps beyond what is offered by these purification kits alone.

how can i efficiently concentrate my dna libraries after overdiluting them?
To efficiently concentrate your DNA libraries after overdiluting, you can use bead purification. Since you've mentioned performing the procedure at a bead:sample ratio of 0.7, ensure that you thoroughly mix the beads before adding them to your sample. Follow the specific steps in Protocol C for Purification of Amplified Libraries starting from step 7. Additionally, if you have any remaining library volume that was not diluted or if you're able to isolate aliquots from the original concentration, consider pooling those as well to recover some of your target material. Bead purification is effective for concentrating and cleaning up DNA libraries by removing excess reagents and unbound fragments while retaining your desired product.

what could be causing the lack of amplification product in my agarose gel?
There are several potential reasons for the lack of amplification product in your agarose gel. First, it's essential to ensure that the primers you designed have a gene-specific melting temperature (Tm) that is close enough—ideally within 4°C of each other—to prevent imbalanced PCR conditions. If there's a significant difference in Tm, it can lead to inefficient amplification. Additionally, if your annealing temperature is too low or the duration too short, this could hinder proper binding of the primers to their target sequences. Consider increasing your annealing time to around 15 seconds and optimizing the overall PCR program according to your specific needs. Another factor could be related to potential repeat sequences where primers may bind non-specifically or amplify shorter unintended products. Blasting your primers against the template DNA can help identify any non-specific binding sites. Lastly, using an appropriate polymerase for your construct size is crucial as well; ensure you're utilizing one suited for amplifying longer fragments if applicable. Reviewing and adjusting these parameters should help improve amplification results. If you're still encountering issues after making these adjustments, consider sharing your primer design file for further analysis and recommendations on possible optimizations.

what's the difference in cDNA product yield between peripheral blood and bone marrow samples?
The cDNA yield can vary significantly between peripheral blood and bone marrow samples due to differences in cell types, cellular composition, and the abundance of mRNA. Bone marrow typically contains a higher concentration of hematopoietic cells, which may lead to a greater overall yield of cDNA when compared to peripheral blood. Additionally, the quality and method used for RNA extraction can also influence the final cDNA product yield. While specific yields may depend on various experimental conditions, it’s generally observed that bone marrow samples provide more robust results for applications like PCR due to their richness in target mRNAs.

hello bakhyt can you provide us with the normalized mouse brain cdna library?
Unfortunately, a normalized mouse brain cDNA library is not available. However, there is the Mate & Plate™ Library - Universal Mouse (Normalized) specifically designed for Yeast Two-Hybrid screening. If you have any further questions or need assistance with something else, feel free to ask!

think thats primers longer try step protocol what temperature should I use for the PCR?
For the PCR protocol using longer primers, it's recommended to use a two-step cycling process. The annealing temperature should be set between 65-68 ºC to ensure proper binding of the primers, particularly for long templates. You can start with an initial denaturation step at 98 ºC, followed by annealing and extension steps at either 68 ºC or slightly lower, depending on your specific primer Tm calculations. Always consider adjusting the extension time based on your target template size; for a 10 kb product, extending the elongation phase may be necessary to ensure complete amplification.

tell storage buffer composition
The storage buffer composition is proprietary, and unfortunately, that information isn't available. If you have any other questions or need further assistance, feel free to ask!

why aren't my primers generating an amplification product?
There could be several reasons why your primers are not generating an amplification product. First, check the melting temperatures (Tm) of your gene-specific primers; if they differ by more than 4°C, it can lead to an imbalanced PCR reaction and hinder product synthesis. Additionally, ensure that you are using a suitable polymerase for the construct size and that your PCR program is optimized. Consider increasing the annealing time in your protocol as well; extending it to around 15 seconds might improve results. If possible, use the primer design tool from Takara Bio to verify Tm values and other parameters related to primer design. If you encounter issues with primer design or need further assistance, sharing the details through their feedback form may provide additional insights. Lastly, providing information about the specific polymerase used and any unique conditions applied during amplification will help diagnose potential issues more accurately.

Can you provide details on potential corporate sponsorship opportunities for the vendor show symposium?
For potential corporate sponsorship opportunities for the vendor show symposium, it's best to reach out directly to the event organizers or the designated contact person associated with the symposium. They can provide specific details on sponsorship levels, benefits, and any relevant deadlines. Additionally, reviewing past events can offer insights into available packages and how companies have participated previously.

what is the shelf life of nucleobond xtra bac kit and how do I check expiration date on reagents?
The shelf life of the NucleoBond® Xtra BAC kit is 39 months from the manufacturing date. To check the expiration date on reagents, you can look for a printed expiration date or batch number on the packaging. This information will typically be located near the barcode or label and may indicate when the product was manufactured and its corresponding expiry period based on its shelf life. If you're unsure, consulting product documentation or contacting customer support can also provide clarity.

where can i find ultra low input rna kit for bacterial rna?
You can find ultra low input RNA kits for bacterial RNA from various scientific supply companies. Popular suppliers include major life science brands such as Thermo Fisher Scientific, Qiagen, and Bio-Rad. Additionally, checking online marketplaces or academic suppliers might yield options that suit your needs. Always ensure to review product specifications to confirm compatibility with your specific requirements before making a purchase.

what are optimal spin times and speeds for transducing cells with retronectin-coated plates?
The optimal spin times and speeds for transducing cells with retronectin-coated plates can vary depending on the specific cell line being used, as well as other experimental conditions. It's generally recommended to start with a range of centrifugation settings—often around 800 to 1200 x g for 30 minutes—as these are common parameters in many studies. However, since each cell line may respond differently, it's beneficial to conduct preliminary tests to determine the best conditions for your particular setup. Additionally, if you have access to data from previous experiments or publications that utilized similar methodologies, reviewing those could provide further insights into effective spin times and speeds. If you're looking for more precise recommendations or data collection regarding this topic, it might be helpful to reach out to experts in your lab or consult relevant literature focused on T-cell transduction using retronectin.

what are the benefits of using guanidinium for small-scale tests without needing to dialyze before sds-page?
Using guanidinium hydrochloride (GuHCl) for small-scale tests offers several benefits without the need to dialyze prior to SDS-PAGE. Firstly, GuHCl is a strong denaturant that effectively unfolds proteins, allowing for more accurate assessment of their molecular weight and structure during SDS-PAGE. This can be particularly useful in determining protein purity or identifying degradation products. Additionally, GuHCl solubilizes many proteins that may otherwise aggregate or precipitate under non-denaturing conditions. As a result, it increases the yield of soluble protein available for analysis. Since you don't need to perform dialysis before running SDS-PAGE, this approach saves time and resources in sample preparation. Furthermore, using GuHCl allows for direct comparison between different proteins or conditions since it provides a consistent environment within your samples without introducing variability from buffer exchange processes. Overall, it's an efficient method for characterizing proteins at smaller scales while maintaining accuracy in results.

how do i calculate the viral gc/ml concentration from my qpcr results using the undiluted supernatant and concentrated stock?
To calculate the viral genome copies per milliliter (gc/ml) concentration from your qPCR results, you can follow these steps: 1. **Determine the qPCR Result**: Record the number of genome copies detected in your undiluted supernatant from your qPCR assay. This value is often given as a copy number per reaction. 2. **Account for Dilution Factor**: If you diluted your samples before qPCR, multiply the detected copy number by the dilution factor to get back to the original concentration in gc/ml. 3. **Volume Considerations**: If you're working with concentrated stock, note how much volume was processed during purification or concentration. You may need to adjust based on any volume changes that occurred during this process. 4. **Calculate Concentration**: - For undiluted samples, if you have calculated a certain number of genome copies per reaction and know how many microliters were used in that reaction, convert it into gc/ml by scaling up according to your total volume. - For example, if you had 1000 gc/reaction and used 5 µl out of a total of 1 ml (1000 µl), then: \[ \text{Total GC} = \left(\frac{1000 \text{ gc}}{\text{reaction}}\right) \times \left(\frac{1000}{5}\right) = 200000 \text{ gc/ml} \] 5. **Adjust for Purification Yield**: Since no purification method is perfect (e.g., assuming 75% yield), consider adjusting downwards accordingly if needed based on expected losses during purification. 6. **Consider Free Genomes Losses**: Remember that free genomes may not concentrate effectively through certain methods; thus, comparing treated and untreated cultures could lead to different counts—untreated might show higher overall counts due to retained free genomes while treated focuses more on functional titers. By following these calculations and considerations, you'll be able to accurately determine the viral gc/ml concentrations from your experimental data!

hello please provide contact information requesting verification employment yes email address fax number send
To request employment verification for Takara Bio USA, you may need to contact their HR department directly. Since there is currently no one available in HR to provide immediate assistance, it's advisable to leave your telephone number so they can reach out to you once someone is available. Unfortunately, specific contact details such as email addresses or fax numbers are not provided at this time. It would be best to follow up with them later for the necessary information.

am wondering can you confirm if using product for internal research will pose any legal or licensing issues?
Using the product for internal research should generally not pose any legal or licensing issues, as it is offered for research use only to both biotech/pharma and academic entities. However, if there are plans to use the product for commercial purposes, a license may be required. It's always a good idea to check specific details with customer service to ensure compliance with all applicable regulations and requirements.

do you sell elution columns and buffer columns separately?
Yes, elution columns and buffer columns are available for purchase separately. The NucleoBond® Xtra Maxi and Midi Columns can be acquired independently, while the EF-grade buffers are also sold separately. You can check the specific product listings on the Takara Bio website for more details and pricing options.

am wondering if there are any restrictions on using these products for internal research purposes?
There are no specific restrictions on using these products for internal research purposes in biotech, pharma, or academic settings. However, it's important to note that if you intend to use the products for commercial purposes, a license may be required. For any further inquiries or clarification regarding product usage and availability, reaching out to customer service is recommended.

what is the process for using g-block block?
To use a gBlock for cloning, you'll want to ensure that your gBlock includes the necessary flanking sequences that overlap with the ends of your linear vector. This is crucial for successful insertion during the cloning process. 1. **Designing Your gBlock**: Use a primer design tool, like the one available on the Takara site, to confirm and add any required sequences at both ends of your gBlock. These additional sequences should be complementary to the regions in your linearized vector where you plan to clone. 2. **Linearizing Your Vector**: Prepare your linear vector by cutting it with appropriate restriction enzymes or using PCR methods if you're generating blunt ends. 3. **In-Fusion Cloning**: When performing In-Fusion cloning, you'll need those overlapping sequences at both ends of your gBlock so they can anneal properly with the corresponding regions on the linearized vector during transformation. 4. **Transformation and Screening**: After mixing together your prepared vector and gBlock insert, follow standard procedures for transformation into competent cells and screen colonies for successful integration of your insert. For more detailed guidance on this process, including visual aids and explanations of how In-Fusion works, consider checking out videos linked on Takara’s learning center page regarding In-Fusion Cloning applications and mechanisms.

can you recommend a company that sells bacillus expression vectors?
There are several companies that offer Bacillus expression vectors. One well-known option is Addgene, which provides a variety of plasmids and vectors for different applications, including those suitable for Bacillus subtilis. Additionally, companies like New England Biolabs (NEB) and Thermo Fisher Scientific also have resources and products related to Bacillus expression systems. It's advisable to review their catalogs or websites for specific options that meet your needs.

how can I successfully amplify a specific region of my non-linearized plasmid without including the unwanted parts?
To successfully amplify a specific region of your non-linearized plasmid while excluding unwanted sequences, you can follow these steps: 1. **Design Primers**: Use a reliable primer design tool, like the one offered by Takara. Input your plasmid sequence and specify the target region you want to amplify. The tool will help generate suitable primers. 2. **Confirm Primer Specificity**: Ensure that the designed primers specifically bind to your desired target without mismatches in the unwanted regions. You may want to check for secondary structures or potential dimerization. 3. **Optimize PCR Conditions**: Adjust PCR conditions including annealing temperature and extension time based on your primer characteristics and template complexity to enhance specificity. 4. **In-Fusion Cloning (Optional)**: If necessary, use In-Fusion cloning technology which allows for seamless assembly of your amplified product while enabling selective deletion of unwanted sequences if they are included during amplification. 5. **Verify Amplification Product**: After performing PCR, analyze the products via gel electrophoresis to confirm the correct size of amplification corresponding only to your desired sequence. By following these guidelines, you should be able to efficiently amplify just the specific region you're interested in from your plasmid without incorporating any undesirable parts.

how long need wait for temporary password to place order?
The temporary password process can vary depending on the account type. For a research account, it may be resolved the same day, while for a purchasing account, it could take a couple of days due to necessary document exchanges. If you haven't received your temporary password yet, it's a good idea to check your junk folder in case the email was misdirected.

how do restriction enzymes function in the presence of Taq buffer?
Restriction enzymes function by recognizing specific DNA sequences and cutting the DNA at these sites. The activity of restriction enzymes can be influenced by various factors, including the buffer in which they are used. Taq buffer is typically designed to support the activity of Taq polymerase during PCR reactions, providing optimal conditions for enzyme stability and functioning. However, when using restriction enzymes in a Taq buffer environment, it’s important to consider that not all buffers are compatible with every enzyme. Some restriction enzymes may require different ionic strengths or pH levels than what is provided by Taq buffer. It's essential to check the manufacturer's recommendations for both the restriction enzyme and any additives (like salts) present in your reaction mix. If compatibility issues arise, you could either switch to a more suitable buffer specifically formulated for use with restriction enzymes or adjust the components accordingly while ensuring that all necessary conditions—such as temperature and time—are optimized for effective digestion of DNA.

what could be the possible reasons for my pcr not working?
There could be several reasons for your PCR not working. Some potential issues include: 1. **Secondary Structures**: If there are secondary structures in the template DNA or primers, they may hinder amplification. A touch-down protocol might help alleviate this issue. 2. **Primer Design**: Ensure that your primers have an appropriate melting temperature (Tm) and that they do not form dimers or secondary structures themselves. 3. **GC Content**: High GC content can lead to difficulties in denaturation and extension phases of PCR, which might require adjustments such as adding DMSO to the reaction mix. 4. **Template Quality & Quantity**: The amount and quality of plasmid DNA used can significantly impact PCR efficiency. Make sure you’re using sufficient high-quality material suitable for amplification. 5. **Cycling Conditions**: Verify that your cycling conditions are optimized for the specific reactions you're performing, including annealing temperatures and extension times based on amplicon length. 6. **Reagents Quality**: Using expired or improperly stored reagents can also lead to poor results. If you've already checked these factors but still face challenges, it may be helpful to consider running a control reaction with known successful conditions to narrow down the problem further.

need confirm if dnase treatment sufficient before rna extraction or should use column for dna removal?
For RNA extraction, using DNase treatment is generally sufficient to remove genomic DNA contamination. However, it's important to ensure that the DNA removal column is not overloaded during this process. If you're working with a high cell count or suspect significant gDNA contamination, you might want to consider both DNase treatment and utilizing the DNA removal column for optimal results. Following the updated protocol will also help ensure that you achieve the best outcomes in your application.

can you provide a detailed protocol for site-directed mutagenesis using the Takara primer design tool and Snap Assembly Master Mix?
For site-directed mutagenesis using the Takara primer design tool and Snap Assembly Master Mix, follow these general steps: 1. **Design Primers**: - Use the Takara Primer Design Tool to create primers that introduce the desired mutation(s). Ensure that your primers are designed to have similar melting temperatures (Tm) and include 15-30 base pairs on either side of the mutation. 2. **Prepare PCR Reaction**: - Set up a PCR reaction with your plasmid DNA as a template (typically between 100 pg to 1 ng). - Include high-fidelity DNA polymerase suitable for amplification. - Add dNTPs, buffer, and your designed primers into the reaction mix according to manufacturer's instructions. 3. **Run PCR Amplification**: - Perform thermal cycling according to your polymerase's recommended parameters (usually involves denaturation, annealing, and extension steps). 4. **Purify PCR Product**: - Purify the amplified product using a gel extraction kit or spin column purification method to remove excess primers and dNTPs. 5. **Remove Template DNA (if necessary)**: - If you used a higher amount of template in step 2, consider digesting any leftover template DNA using Cloning Enhancer from Takara Bio following their protocol guidelines. 6. **Set Up In-Fusion Recombination Reaction**: - Combine purified mutagenized PCR product with linearized vector in accordance with Snap Assembly Master Mix instructions. - Incubate at room temperature for approximately 15 minutes for efficient recombination. 7. **Transform Competent Cells**: - Introduce the recombined vector into competent cells through heat shock or electroporation depending on cell type. 8. **Recovery Culture**: - Allow cells to recover in SOC medium or LB broth without antibiotic selection for about an hour before plating onto selective media containing appropriate antibiotics. 9. **Screen Colonies**: - After incubation, screen colonies via colony PCR or sequencing methods to confirm successful incorporation of mutations. This is a generalized protocol; specific details may vary based on experimental conditions or particular reagents used from Takara Bio products like Snap Assembly Master Mix and Cloning Enhancer. Always refer back to manufacturer protocols for precise procedures tailored towards their products when available.

what's best kit isolating measles rubella rna for downstream rt-qpcr?
For isolating RNA from measles and rubella viruses for downstream RT-qPCR, the NucleoSpin® RNA Virus kit would be a suitable choice. This kit is designed to efficiently isolate viral RNA from various samples, including nasopharyngeal swabs. It can handle sample volumes of up to 150 µl, making it ideal for processing your specimens. Additionally, ensure that you follow the manufacturer's protocols closely for optimal results in your RT-qPCR analysis. If you're looking for alternatives or specific performance comparisons, consider checking other kits specifically validated for these viruses as well.

what are technical benefits differences between using hygromycin selection marker and another antibiotic selection marker for my vector?
Using the hygromycin selection marker offers several technical benefits compared to other antibiotic selection markers. One key advantage is its effectiveness in various cell types, including mammalian cells, which can facilitate successful integration and expression of your gene of interest. Hygromycin also has a distinct mode of action—by inhibiting protein synthesis—that may provide more reliable selection under certain conditions. Another benefit is that hygromycin resistance genes are often less prone to cross-resistance with other antibiotics, making it easier to select for transformed cells without affecting untransformed ones. Additionally, hygromycin can enable higher selective pressure during the screening process, which helps ensure that only successfully transformed cells survive. Choosing between hygromycin and another antibiotic marker ultimately depends on your specific experimental needs and the characteristics of the host organism you are working with. Each antibiotic may have unique properties that could be advantageous depending on factors such as growth rates or ease of use in your particular system.

why isn't the infusion working for inserting the guiderna construct?
The In-Fusion cloning may not be working for inserting the gRNA construct due to several factors. Firstly, if the guide sequence being added is too short or lacks sufficient homology with the vector, it can lead to low efficiency in recombination. The recommendation of adding more than just 20 bp on each side of the insert could help improve this efficiency. Additionally, ensure that your primers are designed correctly and include necessary sequences for linearizing the vector while also incorporating overlaps that facilitate In-Fusion assembly. If there are any mismatches or errors in primer design, it could hinder successful insertion. Another consideration is whether the reaction conditions (like temperature and enzyme concentrations) are optimized according to protocol guidelines, as suboptimal conditions can affect enzyme activity and overall cloning success. Lastly, examining other potential issues such as DNA purity or quantity might be worthwhile—contaminated or degraded DNA can impact results significantly. Reviewing these elements should help identify why In-Fusion isn't yielding desired results for your gRNA construct insertion.

what is the total allowed cycling time for master mix storage and how many freeze-thaw cycles are permitted?
The total allowed cycling time for the master mix storage is approximately one hour, considering 10 to 14 cycles plus a 2-minute initial melt. It’s recommended to store the reaction mix at -20°C for stability, especially if it contains glycerol. While there isn’t a specific limit mentioned on freeze-thaw cycles, it's best practice to minimize them as much as possible to maintain the integrity of the reagents. If you plan to use all reactions within a day or so, storing in a fridge could be acceptable, but long-term storage at -20°C is preferred.

know think batch not good clearly recombining secquence utrs exactly reca cells?
It seems that you are concerned about potential recombination issues with the Stellar competent cells, especially given their recA1 mutation and your specific construct containing repeat sequences. While it’s true that recA1 mutants generally have reduced homologous recombination capabilities, there can still be occasional events depending on various factors such as growth conditions or cell batch variability. If you suspect that the current batch is problematic due to observed recombination with UTRs, it's a good idea to try a different lot of Stellar cells for comparison. The technical team has offered to process a replacement for you once they receive the necessary order information. This could help clarify whether the issue is related to this specific batch of competent cells.

what suitable dna polymerase would you recommend for pcr with fluorescent-tagged primers?
For PCR with fluorescent-tagged primers, it would be advisable to use Ex Taq DNA Polymerase from TaKaRa. This polymerase is known for its tolerance to modified dNTPs, making it a suitable choice for your needs. It's also beneficial to stick with the same conditions that previously worked well for you when using this enzyme. Avoid HiFi enzymes due to their 3' exonuclease activity, which may not be compatible with your setup.

What is the status of my order?
It seems there was a misunderstanding in the conversation, as it focused on preparing a wash buffer rather than discussing an order status. For information regarding your order, it's best to check your email confirmation or contact customer support directly for updates. They should be able to provide you with the most accurate and timely information regarding your order status.

is there a way to confirm if the issues I'm encountering with the in-vitro transcription and extraction protocols are due to component stability or my sample quality?
To determine if the issues with your in-vitro transcription and extraction protocols are related to component stability or sample quality, consider a few approaches. First, verify the expiration date of all components you are using; as confirmed, the lot number for your Guide-it™ In Vitro Transcription Buffer is valid until November 21, 2025. Ensure that all reagents have been stored under appropriate conditions. Next, assess the yield of your guideRNA after purification. If yields are low despite following proper protocols and washing steps thoroughly, it may indicate an issue with either reagent stability or sample quality. Additionally, review your sample handling procedures. For instance, ensure that no wash buffer remains in tubes before eluting RNA and consider allowing silica to dry slightly before elution to improve yield. If possible, conduct a side-by-side comparison by using fresh reagents or samples known to perform well against those currently being used. This can help isolate whether it's a problem with specific components or inherent issues with the samples themselves. Monitoring these factors should provide clearer insights into where any problems might lie.

where is my tracking number?
Your tracking number is 284548147501. You can track your shipment using this link: [FedEx Tracking](https://www.fedex.com/fedextrack/?trknbr=284548147501&trkqual=2460696000~284548147501~FX). The product was shipped on Monday and delivered on Tuesday, signed for by Z.Dan at 12:19 PM. If you need further assistance, feel free to ask!

is it possible to perform single base deletion using mutagenesis with primers that skip a nucleotide?
Yes, it is possible to perform single base deletion using mutagenesis with primers designed to skip a nucleotide. The primer design tool can help create these specific primers for your PCR reaction, which will allow you to delete the target base and then recircularize the vector following the In-Fusion reaction. For best results, ensure that you're using a high-fidelity polymerase like PrimeStar Max, especially if you're working with larger plasmids up to 15 kb.

are there any recommended adapters for ligation-based workflows with t-base overhangs?
For ligation-based workflows utilizing T-base overhangs, it's important to choose adapters that are specifically designed for this purpose. While the provided link focuses on unique dual-index kits compatible with NGS applications, you may want to explore other products or manufacturers that offer adapters tailored for T-overhang compatibility in your specific workflow. Checking with suppliers like Takara Bio or similar companies could yield suitable options that meet your requirements for ligation-based methodologies.

is there a cost-effective option for introducing single base replacements in pcdna plasmids?
A cost-effective option for introducing single base replacements in pcDNA plasmids is to use site-directed mutagenesis techniques. One widely used approach is the PCR-based method, which allows for specific alterations in the DNA sequence using primers that incorporate the desired mutations. Kits designed specifically for site-directed mutagenesis are often available and can be more economical than purchasing multiple components separately. Additionally, consider checking if any bulk purchase options or discounts are offered by suppliers for kits that include necessary reagents like polymerases and competent cells, as this can further reduce costs while providing all required materials for your experiments.

what should we do about the lentix concentrator fridge breakdown and its impact on virus concentration?
In the event of a breakdown of the lentix concentrator fridge, it is crucial to assess the status of any samples stored inside. If they have been compromised or allowed to evaporate due to temperature fluctuations, it's important to evaluate their viability for virus concentration work. To mitigate the impact on virus concentration, consider these steps: 1. **Inventory Check**: Review all stored materials and determine which samples were affected. 2. **Temperature Monitoring**: Confirm how long and at what temperatures samples were exposed during the breakdown. 3. **Consultation with Experts**: Engage with your team or specialists in virology for insights on whether any remaining samples can still be used effectively based on their stability and integrity after exposure. 4. **Replacement Options**: Explore options for replacing lost materials if necessary and prioritize repairs or replacements for the fridge promptly. 5. **Preventive Measures**: Implement protocols to monitor equipment more closely moving forward, ensuring backup systems are available in case of future issues. Taking these actions will help ensure minimal disruption in your research efforts while maintaining sample integrity as much as possible.

is it safe to use bleach around lenti-x lenti concentrator during virus concentration?
Using bleach around a Lenti-X Lenti concentrator during virus concentration is not recommended. Bleach can produce harmful fumes and may interfere with the equipment or samples. It's important to follow the manufacturer's guidelines for cleaning and disinfecting in laboratory settings. Consider using alternative disinfectants that are safe for use in biosafety contexts, ensuring proper ventilation and safety protocols are maintained.

what is the best way to resolve my issue?
To resolve your issue effectively, consider analyzing the search results on Google Scholar for relevant publications related to "smarter race 5 3 kit." Review the titles and abstracts to identify which papers are most pertinent to your needs. If you're looking for specific information or solutions outlined in those publications, you may want to access them directly through institutional resources or request copies from authors if necessary. Additionally, engaging with online forums or communities focused on this topic could provide further insights and assistance.

Does the ssRNA ladder marker contain formaldehyde, and is it necessary to use a loading buffer with the product?
The ssRNA ladder marker typically does not contain formaldehyde; however, it's important to verify the specific formulation from the manufacturer since variations can exist. Regarding the use of a loading buffer, it is essential for ensuring proper denaturation of RNA and preventing secondary structures during electrophoresis. A loading buffer helps maintain an appropriate environment for visualization and effective separation on a gel. Therefore, using a suitable loading buffer with your RNA sample is recommended to achieve optimal results in your experiments.

could the issue be related to the new plasmid construction or the sd leu plates not being properly tested for leucine control?
Based on the conversation, it seems that the primary concern is with the transformation process rather than the plasmid construction or the sd leu plates. If control plasmids are not yielding any colonies, it suggests that there may be an issue with either the transformation procedure itself or potentially with media quality. It's important to ensure that all components used in the experiment are within their expiration dates and functioning properly. If you suspect issues with leucine control on sd leu plates, testing those plates separately could be beneficial. Additionally, verifying whether proper protocols were followed during both plasmid preparation and yeast transformation can help identify where things might have gone wrong. Overall, focusing on troubleshooting the transformation process and confirming media viability would likely provide clearer insights into your current challenges.

aav purification method different than mlv?
Yes, the purification methods for AAV (Adeno-Associated Virus) and MLV (Murine Leukemia Virus) are different due to their distinct properties and requirements. The AAVpro Concentrator is specifically designed to purify and concentrate AAV from viral supernatants, while the Retro-X concentrates work primarily with retroviruses like MLV. Each method may require specific protocols tailored to the unique characteristics of the respective virus type for effective purification. If you need more detailed information about these methods or have further questions, feel free to ask!

am unable connect anyone either phone email
It seems that there have been some delays in communication, and it can be frustrating when you're unable to connect with someone for assistance. If you’re looking for support, consider checking the official website or help center of the organization for alternative contact methods or resources. They may also provide a live chat option or additional email addresses to reach out to. Your patience is appreciated, and hopefully, you'll receive the assistance you need soon.

How can I optimize lysis for small sample sizes in Smart-seq library prep, specifically for blastocyst embryos?
Optimizing lysis for small sample sizes, such as blastocyst embryos, in Smart-seq library preparation can be challenging since this kit has not been tested specifically with embryos or blastocysts. However, here are a few strategies that may help: 1. **Minimize Lysis Volume**: Use the smallest effective volume of lysis buffer to ensure that you are working with concentrated samples. 2. **Increase Incubation Time**: Consider prolonging the incubation time during the lysis step to allow for more complete cell disruption while being cautious of potential RNA degradation. 3. **Use Gentle Lysis Methods**: Instead of using harsh chemical methods, you might explore using mechanical disruption techniques like pipetting up and down gently or using a microcentrifuge tube pestle to break open cells without damaging RNA. 4. **Optimize Temperature Conditions**: Perform lysis at lower temperatures if possible to reduce RNA degradation and maintain integrity during processing. 5. **Pilot Experiments**: Conduct preliminary experiments with known quantities of material similar in size and structure to your target samples (e.g., smaller cell lines) before applying these protocols directly on blastocysts. 6. **RNA Stabilization Solutions**: Adding an RNA stabilization solution immediately after lysis could help preserve the quality of isolated RNA from sensitive samples like embryos. It's essential to validate any modifications through controls when transitioning from suspension cells used in previous validations with this kit, ensuring reliable results for your specific application.

what do i need to do to fix the scanning issue?
To address the scanning issue, first ensure that the scanner is properly connected to your computer and powered on. Check for any error messages on the scanner's display or your computer screen. If there are no visible issues, try restarting both the scanner and your computer. Additionally, verify that the correct drivers are installed for your scanner model and consider updating them if necessary. If problems persist, consult the user manual for troubleshooting steps specific to your device or reach out to technical support for assistance.

what's the best product for reliable mouse genotyping?
For reliable mouse genotyping, the Terra PCR Direct Polymerase Mix is a strong option, especially for working with whole or crude tissue extracts. It simplifies the workflow and is designed to handle direct PCR applications effectively. Additionally, you may consider using the NucleoSpin Tissue kits for genomic DNA purification from various tissue samples, including mouse tails. When choosing primers for your reaction, it's important to note that in the SMARTer smRNA-Seq Kit (635029), you can utilize F2 forward primer along with R1-R12 reverse primers. For 635031, both F1-F4 forward primers and R1-R12 reverse primers are applicable. While there are alternative reagents available that could mimic this genotyping kit's performance when used together with Terra polymerase, opting for dedicated products like these can save time and effort in optimizing your assays. Overall, while no product is universally perfect due to varying assay designs and requirements, Terra offers good reliability in routine single target scenarios within typical amplicon sizes.

increase magnet time for isopure mini extract, what settings should I use?
To adjust the magnet time for the IsoPure Mini extraction, it's important to ensure that you are following any specific guidelines provided in the protocol. Since it appears that the protocol has already been optimized by Macherey Nagel's automation team and there is no additional wash step included, consider maintaining consistency with their established settings. If you're looking to increase the magnet time beyond what is suggested in your current protocol, a good starting point would be to incrementally test longer durations while monitoring outcomes to find an optimal balance without compromising efficiency or yield. Additionally, ensuring proper alignment of your tubes with the magnets can enhance performance. For tailored advice or adjustments based on your specific setup and issues faced during processing, reaching out directly to Macherey Nagel might provide you with more precise recommendations.

what is the difference in activity between cold-active Cryonase and regular Benzonase at low temperatures, specifically regarding RNA cleavage?
The primary difference in activity between cold-active Cryonase and regular Benzonase at low temperatures lies in their enzymatic efficiency. Cold-active Cryonase is designed to maintain higher enzymatic activity at lower temperatures, which allows for effective RNA cleavage even when the reaction conditions are not optimal, such as during cold storage or specific experimental setups that require reduced temperatures. In contrast, regular Benzonase typically exhibits decreased activity at low temperatures, making it less efficient for RNA cleavage under similar conditions. This characteristic of Cryonase can be particularly advantageous in applications where maintaining nucleic acid integrity while working with temperature-sensitive samples is crucial. For a more detailed comparison on specific activities and substrate preferences, including whether either enzyme acts as a 5’, 3’, or both exonucleases, consulting product literature or technical support from the manufacturer would provide valuable insights.

what do i do next with the colonies?
Next, you should pick one or two colonies from your cloning project and culture them. After that, perform a sequencing check to verify if the insert is correctly cloned into your vector. It’s also important to ensure that both the linearization of your vector and PCR reactions for the vector and insert went well with the expected sizes. If everything checks out, you can proceed based on those results. If any issues arise during sequencing, consider reviewing your design using the available tools provided by the kit manufacturer.

think need optimize qpcr product for cloning?
Optimizing your qPCR product for cloning can be beneficial, especially if you're working with short fragments like 100-200 bp. Since qPCR buffers are specifically designed for shorter DNA products, they may yield better results in terms of efficiency and quality compared to standard PCR protocols. You might consider using a protocol that involves adding an A overhang by incubating the product at 72 degrees after amplification. This method is commonly used for TA-cloning and could enhance your cloning success rate. If you have specific concerns about yield or other aspects of your experiment, it would be great to look into alternative enzymes or conditions as well. If you need further assistance or specific guidance on optimizing your conditions, feel free to ask!

how can i improve virus recovery and stability during the lenti-x concentration process?
To enhance virus recovery and stability during the Lenti-X concentration process, consider the following strategies: 1. **Optimize Incubation Time**: While longer incubation may lead to more precipitated virus, it's essential to balance this with diminishing returns after a certain point. Stick closely to recommended incubation times for optimal results. 2. **Use Serum-Free Medium**: If possible, use a serum-free medium for your viral cultures or concentrate conditions. This can reduce the co-precipitation of serum proteins that may complicate recovery. 3. **Centrifugation Technique**: Ensure you employ adequate centrifugation speeds and durations when pelleting the virus post-precipitation. Natural sedimentation alone is insufficient; proper centrifugation will help obtain a cleaner pellet. 4. **Dissolving Pellets**: Be prepared that some serum proteins might not dissolve easily post-centrifugation, but typically, virus particles should re-suspend well in solution. You can discard any undissolved protein while retaining dissolved components that generally do not impede infection. 5. **Avoid Prolonged Storage of Pellets**: Minimize time between collection and processing of pellets to maintain viral integrity. 6. **Monitor Stability Conditions**: Keep viruses under appropriate temperature and pH conditions immediately after concentration to preserve their infectivity throughout handling processes. By implementing these tips, you should see an improvement in both recovery rates and overall stability during your lenti-x concentration process.

what is the required read length for smart-seq when using one end indexed with dual indexing?
The recommended read length for SMART-Seq when using single-end sequencing is 50 base pairs (bp). It’s important to note that while dual indexing can be applied in this context, the primary focus should remain on utilizing single-end reads with the appropriate custom primer, particularly due to considerations related to low diversity A/T regions. For further details regarding your specific instrument and any necessary adjustments, refer to the manufacturer's guidelines.

what is the preferred product for drop dialysis purification?
The preferred product for drop dialysis purification is the Nucleospin gDNA Clean-Up XS. This method offers a more efficient and safer alternative compared to traditional techniques, which can risk sample loss if not handled carefully. The link to the product provides further details on its application and benefits for DNA purification.

why won't mn beads work instead of ampurexp?
The reason MN beads are not recommended as a substitute for Ampure beads is that there hasn't been internal testing or data to support their effectiveness in this specific procedure. Without established results, it's difficult to ensure they will perform adequately. Therefore, it would be safer for customers to continue using Ampure beads unless they are willing to experiment and invest the necessary time and effort into determining how MN beads might work in their particular application.

I haven't received my research account yet; can you help me with that?
For assistance with your research account, it's best to reach out directly to customer support. You can call the orders line for help or send an email to the appropriate support address provided by the company. They will be able to assist you with any issues regarding your account.

how can i keep huvec cells adherent without causing sponge to dissolve?
To keep Huvec cells adherent without causing the sponge to dissolve, it's important to carefully manage the conditions during and after application. First, ensure that EDTA is maintained in the cell suspension until just before applying it to the sponge; this will help keep the cells in suspension without allowing them to attach prematurely. After applying the cell suspension onto the sponge, you can gently spin down any excess liquid and discard it while making sure not to disturb the adhered cells. If needed, adding a wash step could help remove excess EDTA or trypsin while minimizing any impact on cell adhesion. It's also crucial to choose a sponge material that is compatible with your specific type of Huvecs and allows for their adherence without compromising its structural integrity. Experimenting with different times of exposure and handling techniques may provide further insights into optimizing adherence while preserving both cell viability and sponge functionality.

what is the maximum size of plasmid that your kit can purify?
The maximum size of plasmid that the kit can purify typically varies by manufacturer, but many kits are designed to handle plasmids up to 25 kb in size. It's always best to consult the specific product documentation for precise information regarding your particular kit.

cdna synthesis successful or not?
The success of cDNA synthesis can be influenced by several factors, including the presence of PCR inhibitors and the quality of the starting material. If there is no amplification detected during PCR, it may indicate that cDNA synthesis was not successful or that there are issues such as low template concentration or inhibition from contaminants. The recommended 260/230 ratio should ideally be higher than 1.6 to confirm good purity, as this could affect the efficiency of cDNA synthesis. Diluting your cDNA sample 10-100 fold prior to PCR can also help improve results by reducing potential inhibitors. Checking for specific known inhibitors in your samples can provide further insights into any challenges you might face with amplification.

what troubleshooting recommendations do you have for smrna seq kit since none of my samples worked but the positive control did?
When troubleshooting issues with an sRNA sequencing kit where the positive control works but your samples do not, consider the following recommendations: 1. **Sample Quality**: Ensure that your RNA samples are of high quality. Check for degradation using a bioanalyzer or similar method to assess integrity. 2. **Input Concentration**: Verify that you are using the correct input concentration as specified in the protocol. Too low or too high concentrations can impact performance. 3. **Contamination**: Assess your samples for potential contamination with inhibitors, which could affect amplification and sequencing results. 4. **Reagent Handling**: Double-check that reagents were prepared and stored correctly according to guidelines, and ensure they have not expired. 5. **Protocol Adherence**: Review each step of the protocol carefully to confirm adherence to recommended procedures without deviations. 6. **Reaction Conditions**: Make sure all reaction conditions (temperature, time) were followed accurately during library preparation. 7. **Equipment Calibration**: Ensure that any equipment used (like pipettes or thermocyclers) is properly calibrated and functioning as expected. 8. **Replicates Testing**: If possible, run additional replicates of your samples alongside controls to determine if there is consistent failure across multiple tests or isolated issues with specific samples. If these steps do not resolve the issue, it may be beneficial to consult technical support directly for further assistance tailored specifically to your situation.

how do i properly perform in-fusion cloning with my pcr amplified fragments to ensure correct fusion of the three dna pieces?
To perform In-Fusion cloning with your PCR amplified fragments and ensure the correct fusion of your three DNA pieces, follow these steps: 1. **Design Overlapping Fragments**: Ensure that each of your PCR products has overlapping sequences at their ends. The overlaps should typically be around 15-20 bp to facilitate efficient joining. 2. **PCR Amplification**: Use flanking primers for amplifying each fragment separately. Avoid including additional sequences in the In-Fusion mix as they can hinder efficiency. 3. **Purify PCR Products**: After amplification, purify your PCR products to remove any residual primers or nucleotides that could interfere with the In-Fusion reaction. 4. **Set Up the In-Fusion Reaction**: Mix equal molar amounts of your purified PCR products according to the manufacturer's guidelines for In-Fusion cloning. This will allow for proper annealing between overlapping regions. 5. **Transformation and Screening**: Transform competent cells with the ligation mixture and plate on selective media to screen for successful clones containing all three fused fragments. 6. **Verification**: Once you obtain colonies, verify the presence of correct constructs through colony screening methods such as restriction enzyme digestion or sequencing. For specific optimization techniques related to overlap extension PCR or further details about improving efficiency during this process, consider reviewing relevant literature such as protocols found in scientific articles focused on transgene construction methodologies. If you have more questions later on, feel free to reach out for support when needed!

know if retro-x and retro go can be mixed with retro supe for a weekend use?
Mixing Retro-X and Retro Go with Retro Supe for weekend use should generally be fine, as long as you monitor the incubation conditions. It's important to ensure that all components are compatible and maintain appropriate storage temperatures. If you're planning to incubate for an extended period, avoid exceeding 3 days at 4°C to prevent potential decreases in viral titer. Always consult product guidelines or your supplier for specific recommendations regarding mixing different reagents.

what are the key differences between the new smart-seq total rna pico input zapr mammalian kit and the older version in terms of ribo depletion, unique dual indexes, and input requirements?
The new SMART-Seq® Total RNA Pico Input (ZapR™ Mammalian) kit offers several improvements over the older version, SMARTer® Stranded Total RNA-Seq Kit v2 - Pico Input Mammalian: 1. **Ribosomal RNA Depletion**: The updated kit features efficient depletion of rRNA-derived cDNAs across a wide input range. It has been specifically noted for its effective treatment with the ZapR Mammalian rRNA Depletion Kit, which enhances the quality and relevance of reads from both low (250 pg) and high (10 ng) total RNA inputs. 2. **Unique Dual Indexes**: In both kits, Unique Dual Indexes must be purchased separately. This remains consistent between the new and older versions. 3. **Input Requirements**: The input requirements differ significantly; the new kit is compatible with 250 pg to 10 ng of total RNA, while the previous version required a minimum of 500 pg to 10 ng for optimal results. These enhancements make the newer kit more versatile in handling lower amounts of starting material while improving overall data quality through better rRNA depletion strategies.

where can i find lentiviral particles for myc overexpression that are not part of an inducible system?
To find lentiviral particles specifically for Myc overexpression that are not part of an inducible system, it would be best to check with suppliers who offer standalone viral vectors. While this conversation indicated that the current offerings may not include a Myc-only lentiviral particle in the catalog, you might consider reaching out directly to manufacturers or distributors specializing in gene delivery systems. They may have options available or could assist you in custom solutions based on your research needs. Additionally, looking into commercial vendors that provide a variety of viral constructs could yield suitable products for your project.

what options available for improving integration efficiency using your kits?
To improve integration efficiency when using our kits, consider the following options: 1. **Optimized Plasmid Design**: Ensure that your plasmid vector is designed for efficient chromosomal integration. The pAUR101 DNA vector is specifically intended for this purpose in S. cerevisiae, facilitating stable integrations. 2. **Selection Markers**: Utilize effective selection markers such as Aureobasidin A to screen for successfully integrated clones. This antibiotic allows only those yeast cells with successful integration to survive on selective media. 3. **Transformation Protocols**: Follow optimized transformation protocols tailored for yeast, ensuring that conditions promote high-efficiency uptake of the plasmids. 4. **Screening Techniques**: Employ robust screening techniques post-transformation to identify and isolate positive clones more effectively. 5. **Supplementary Media**: Consider using specialized growth media or additives that enhance cell viability and integration success rates during culturing. 6. **Consult Technical Support or Literature**: For specific recommendations based on your experimental setup, consulting technical support or relevant literature may provide additional insights into improving your integration efficiency. If you require further assistance or have specific needs regarding these options, please feel free to reach out!

would these backordered kits still meet our research timelines?
To determine if the backordered Unique Dual Index (UDI) kits will meet your research timelines, it would be best to assess the expected delivery dates for those kits and compare them against your project deadlines. If you have flexibility in your timeline or alternative indexing options available, that could also help keep your research on track. Checking with the supplier for updates on shipping times may provide additional clarity on whether these kits can still align with your planned schedule.

how can i effectively detach well-attached hsc cells without harming them for downstream assays?
To effectively detach well-attached hematopoietic stem cells (HSCs) without causing harm for downstream assays, consider the following steps: 1. **Trypsinization**: Use a diluted trypsin solution, ideally in the range of 0.05% to 0.25%. Longer incubation times at this concentration can help gently detach the cells while minimizing damage. 2. **EDTA Addition**: Incorporating EDTA in your detachment solution may enhance cell release by chelating calcium ions that are important for cell adhesion. 3. **Mechanical Disruption**: If needed, you can gently tap or shake the plate after applying trypsin/EDTA to assist with cell detachment without using harsh mechanical forces that could rupture cells. 4. **Temperature Considerations**: Ensure that your solutions are warmed to room temperature or slightly above before use as warmer temperatures can promote better enzymatic activity and reduce stress on cells. 5. **Coating Concentration Adjustment**: Since retronectin concentrations might be higher than recommended (20-100 µg/ml), ensure you're optimizing coating conditions to improve HSC attachment and facilitate easier recovery during detachment processes. 6. **Gentle Pipetting**: After sufficient incubation with trypsin/EDTA, use a pipette to gently resuspend the cells rather than vigorous shaking which could lead to cell lysis. By implementing these methods, you should be able to achieve effective detachment of HSCs while preserving their viability for subsequent applications.

how ensure maintain functional lentivirus during purification process?
To maintain functional lentivirus during the purification process, it's essential to minimize any potential changes that could affect its viability. Here are some recommendations: 1. **Use Appropriate Concentrators**: While concentrators can concentrate proteins from the media, which may cause irritation at the injection site, it’s important to ensure that they do not retain or adsorb viral particles. Amicon filters typically should not stick to viruses. 2. **Consider Direct Use of Concentrated Samples**: Some customers have successfully used concentrated samples directly without adverse effects. Testing a small batch initially could help determine if this method works for your specific application. 3. **Buffer Exchange Protocols**: If there is any concern regarding impurities or protein concentration affecting functionality, performing a buffer exchange might be beneficial. This step helps remove unwanted components and ensures better conditions for virus stability. 4. **Evaluate Purification Needs**: Assess whether purification methods are necessary for your particular application since those procedures can be labor-intensive and introduce additional risks of losing viral function. 5. **Gather Feedback from Users**: Since you mentioned relying on customer feedback in lieu of in vivo work, continue engaging with users who have experience with similar processes—this can provide valuable insights into maintaining lentivirus functionality post-purification. By integrating these strategies into your workflow, you can enhance the likelihood of preserving lentiviral activity throughout the purification process while also being efficient in resource usage.

what modifications do the oligos require for ligation into the vector?
For successful ligation into a vector, oligos typically require specific modifications. These may include: 1. **Phosphorylation**: The 5' ends of the oligos should be phosphorylated to facilitate ligation by providing a reactive phosphate group. 2. **Overhangs or Cohesive Ends**: Depending on the vector and restriction enzymes used, you may need to add compatible overhang sequences at the ends of your oligos to ensure they can anneal properly with the vector. 3. **Length Considerations**: Ensure that the oligo lengths are suitable for efficient ligation; usually around 20-30 nucleotides is optimal. 4. **Purification**: After synthesis, it's often recommended to purify the oligos before ligation to remove any impurities or unreacted components that could interfere with the process. Referencing guidelines provided by suppliers can offer additional insights tailored specifically for their products and protocols.

can you confirm if using the lenti-x lenti packaging single shot for retrovirus production will yield similar viral titers as the lentivirus approach?
Using the Lenti-X Lenti packaging single shot for retrovirus production may yield different viral titers compared to a lentivirus approach. Lentiviral vectors typically achieve higher titers due to their ability to transduce non-dividing cells and their optimized design, which might not be fully replicated in standard retroviral systems. Additionally, factors such as vector self-inactivation and the presence of specific antibiotics can influence overall titer outcomes. It's important to consider these variables when comparing titers between the two methods.

what is the total time for yeast transformation to be completed?
The total time for yeast transformation can vary depending on the specific protocol and steps involved. Generally, the process includes preparing competent cells, mixing them with DNA, and then incubating them under certain conditions. This typically takes several hours to complete. For a detailed timeline: 1. Preparation of competent yeast cells: approximately 1-2 hours. 2. Transformation procedure (mixing DNA with competent cells and incubation): about 30 minutes to 1 hour. 3. Recovery period after transformation: usually around 1-4 hours, depending on the method used. In summary, you can expect the entire transformation process to take anywhere from approximately 3 to 7 hours in total. It's advisable to check specific protocols provided by your kit or lab for more precise timing based on your experimental setup.

what is the best buffer to use for dissolving sponge protocol?
The best buffer for dissolving in the sponge protocol often depends on the specific cell type being used and their preferences. While there are different options available, a common choice is to use a wash solution that contains cations to maintain cell health, especially during the washing step. If you’re using a release buffer that acts as a chelator (like EDTA), it's important to ensure that your cells can tolerate it without additional cations. If your cells respond well with just EDTA and no cations, then that should work fine as well. It's always good practice to consult with R&D or relevant guidelines for optimal results tailored to your specific application.

Is the sequin spike-in spike product available through Takara USA?
The sequin spike-in spike product does not appear to be available in the Takara USA product database. It was also noted that it is not listed on the Takara Japan product site. While there may be a possibility of checking with colleagues at Takara Japan for availability, even if it is available, it might not be possible to sell it in the US. Further inquiries can be made regarding this product's status.

able provide total number plasmids virus prepared?
The virus is packaged using a total of five plasmids for packaging. For more specific details, you can refer to the information page on the Takara Bio website regarding 4th generation lentiviral packaging.

what additional information can you provide regarding the sensitivity of the sstotalrna single cell kit for rna-seq of evs?
The SMARTer Stranded Total RNA-Seq Kit, particularly in its Pico Input Mammalian version, is designed for high sensitivity and specificity in total RNA sequencing applications. However, when working with extracellular vesicles (EVs), there are important considerations regarding potential contamination and the nature of the RNA being profiled. 1. **Sensitivity to Contaminants**: The kit's extreme sensitivity can be a double-edged sword; while it allows for the detection of low-abundance RNAs, it also raises concerns about incorporating contaminants such as ssDNA that may co-purify during RNA isolation from EVs. This unwanted incorporation can lead to misleading results if non-target sequences are amplified alongside genuine RNA targets. 2. **Input Requirements**: The kit typically requires low input amounts (250 pg - 10 ng total RNA), which is advantageous for working with limited samples like those derived from biofluids or isolated EVs. It facilitates profiling even when starting material is scarce. 3. **Library Construction**: Protocol optimization may be necessary to mitigate the effects of contamination and improve library quality. Using stringent purification steps during RNA extraction and possibly employing additional filtration methods could help minimize undesired nucleic acid species entering downstream applications. 4. **Comparison with Other Technologies**: While this kit excels at capturing mature mRNAs and certain long noncoding RNAs, researchers interested specifically in small RNAs (<100 bp) might consider using dedicated kits like SMARTer smRNA-Seq Kit designed explicitly for enriched small RNAs or total RNA inputs ranging from 1 ng–2 μg. 5. **Application Context**: Recent studies have successfully utilized this technology across diverse biological contexts related to human health conditions—such as cancer diagnostics through plasma cfRNA profiling—indicating robust performance but highlighting variable outcomes based on sample type and integrity. Overall, careful consideration of these factors will enhance experimental design when utilizing the SMARTer Stranded Total RNA-Seq Kit for EV-related analyses, ensuring meaningful insights into the transcriptomic landscape they represent.

what is the minimal recommended titer for lentiviruses when changing media?
The minimal recommended titer for lentiviruses when changing media can vary depending on the specific application and cell type being used. However, a common guideline is to aim for a titer of at least 1 x 10^6 infectious units per milliliter (IU/mL) to ensure effective transduction. It's important to consider that higher titers may be necessary for more challenging cells or applications where efficient gene delivery is critical. Always refer to specific protocols or literature related to your experimental conditions for the best outcome.

wondering when sos shipping will arrive
The shipping for the replacement should be processed today, and if everything goes smoothly, it could ship out today for next-day delivery. Please keep an eye on your tracking information once it's shipped to know when to expect its arrival. If you have any further questions or need assistance later, feel free to reach out through the appropriate support channels.

what is the best method to measure virus production in my experiment using lenti-x and vsv-g?
To effectively measure virus production in your experiment using the Lenti-X system and VSVg, follow these steps: 1. **Obtain a Viral Vector:** Ensure you have a viral vector that contains your gene of interest (GOI). If you're looking to monitor viral transduction, consider using the pLVX-ZsGreen1-N1 vector which includes a green fluorescent protein reporter. 2. **Prepare Packaging Cells:** Use the Lenti-X Expression System (EF1α Version) along with the appropriate packaging cell line, such as 632180 or HeLa cells. This setup will allow for efficient production of lentiviral particles. 3. **Transfect Cells:** Transfect your packaging cells with both the plasmid containing VSVg and your chosen viral vector that carries the GOI. This combination will lead to the expression of necessary proteins for virion assembly and release. 4. **Harvest Virus Particles:** After sufficient incubation time post-transfection, collect the supernatant from your packaging cells. The supernatant will contain lentiviral particles. 5. **Filter Supernatant:** Filter this supernatant to remove any remaining cells or debris before proceeding to infection assays. 6. **Infect Naive Cell Line:** Use this filtered supernatant to infect naive target cell lines where you want to study transduction efficiency or expression of your GOI. 7. **Monitor Reporter Gene Expression:** Finally, assess GFP expression in infected target cells as an indicator of successful virus production and transduction efficacy. This method allows you not only to produce viral particles but also provides a clear way to measure their functionality through reporter gene expression in recipient cells.

anyone assist with downloading shasta sequencing information?
It seems that reaching out to the Field Application Scientist might be your best option for assistance with downloading Shasta sequencing information. They can provide targeted support for your needs. Additionally, if you have further questions or require help later on, consider contacting Technical Support directly for more guidance. If you'd like me to send an email to Technical Support on your behalf, just let me know!

how can I determine if the issue is with amplification or genomic background contamination?
To determine if the issue is with amplification or genomic background contamination, you can follow a few steps. First, check the gel electrophoresis results for any unexpected bands outside of the expected size range. If there are additional bands present, this may indicate genomic contamination. Next, consider your miniprep process; if you did a miniprep, it's more prone to include contaminating DNA compared to midi or maxi preps. Additionally, running controls alongside your sample can help differentiate between true amplification and background noise. Using specific primers that target only your region of interest can also reduce non-specific amplification. If possible, repeating the PCR with a dilution series of your template DNA might help clarify whether you're dealing with high concentrations leading to non-specific products or other issues related to the reaction conditions themselves.

any recommendations for alternative expression systems or competent cells?
For alternative expression systems, you might consider using E. coli strains like BL21(DE3) or Rosetta for protein expression, as they are widely used and well-supported. If you're looking for insect cell systems, the Baculovirus system is a popular choice that can be very effective for expressing larger proteins. As for competent cells, if Brevibacillus has been discontinued and you're having difficulty sourcing alternatives, other options include commercially available competent E. coli strains such as One Shot or Stbl3 from various suppliers. It may also be helpful to check out literature specific to your target protein or application; there could be protocols detailing how to make different bacterial strains competent. If you're open to exploring these suggestions further or need more detailed information on any particular system, reaching out to technical support teams of product providers could yield valuable insights tailored to your research needs.

Can you provide details on how to use retronectin with ultra low adhesion plates?
When using RetroNectin with ultra-low adhesion plates, it's important to note that these plates are designed to inhibit cellular attachment due to their covalently bound hydrogel layer. This feature minimizes protein absorption and may affect the binding efficiency of RetroNectin. To successfully use RetroNectin in conjunction with ultra-low adhesion plates, consider the following steps: 1. **Coating Protocol**: If possible, coat the ultra-low adhesion plate surface with a suitable concentration of RetroNectin prior to cell seeding. This can enhance cell attachment despite the low-adhesion properties. 2. **Cell Seeding**: When adding cells, ensure they are in an appropriate medium that supports their growth without encouraging excessive activation or adherence. 3. **Optimization**: Test different concentrations of RetroNectin and incubation times for optimal results based on your specific cell type and experimental needs. 4. **Monitoring Cell Behavior**: Observe how your cells respond under these conditions; adjustments might be necessary depending on their attachment characteristics and growth patterns. 5. **Consulting Manufacturer Guidelines**: Always refer back to any manufacturer guidelines or protocols provided for both the Ultra-Low Attachment Plates and RetroNectin for best practices tailored specifically for those products. These considerations can help you effectively utilize RetroNectin while working within the constraints of ultra-low adhesion surfaces.

do we need heat inactivated fbs for culturing neuroblastoma and breast cancer cell lines with the tet-on system?
Heat inactivation of FBS is generally not necessary for culturing neuroblastoma and breast cancer cell lines, especially if immune effector activity is not a concern. It may be beneficial to perform a comparison by culturing your cells with both heat-inactivated and non-heat-inactivated FBS to assess any differences in growth or behavior, as heating can potentially reduce the growth-promoting properties of FBS.

is anyone available to take my call?
It appears that you can reach out to a Field Application Scientist if you need assistance, and there is also a Senior Manager who could be contacted. If you're looking for immediate support, it may be best to call the provided contact number. Additionally, if no one is available to take your call at that moment, consider sending an email with your inquiries for further assistance.

reagent stability affect pcr results why weak bands?
Reagent stability can significantly impact PCR results, leading to issues such as weak bands. If reagents are degraded or improperly stored, their effectiveness in amplifying DNA may diminish. This can result in lower yield and visibility of PCR products on a gel. Additionally, factors like the template quality (genomic DNA, cDNA, plasmid) and the number of PCR cycles used also play a crucial role. It's important to ensure that all components are fresh and properly handled to achieve optimal amplification results. To troubleshoot further, examining gel images alongside cycling conditions would be beneficial for identifying specific problems related to reagent performance or experimental setup.

am i getting my order today
It seems that the conversation doesn't provide specific information about an order or its delivery status. To get accurate details regarding your order, it would be best to check with the relevant customer service or tracking system associated with your purchase.

What products do you recommend for sensitive skin?
For sensitive skin, it's important to choose products that are gentle and free from potential irritants. Look for fragrance-free cleansers with soothing ingredients like aloe vera or chamomile. Moisturizers containing hyaluronic acid or ceramides can help retain moisture without causing irritation. For sunscreen, mineral-based options with zinc oxide or titanium dioxide are often better tolerated by sensitive skin types. Additionally, consider patch-testing any new products before full application to ensure compatibility with your skin.

can you provide me with a comparison table for the available versions of picoplex wga technology?
Creating a comparison table for the available versions of Picoplex WGA (Whole Genome Amplification) technology involves outlining key features and specifications for each version. Below is a simplified example: | Feature/Version | Version 1 | Version 2 | Version 3 | |---------------------|----------------|------------------|---------------------| | Amplification Method | Isothermal | PCR-based | Hybrid | | Input DNA Amount | Low (<10 ng) | Medium (10-100 ng)| High (>100 ng) | | Yield | Moderate | High | Very High | | Application Areas | Basic research | Clinical diagnostics, Forensics | Comprehensive genomics, Single-cell analysis | | Time to Results | 2 hours | 4 hours | 6 hours | | Compatibility | Various samples.| Specific types | Broad compatibility | This table gives an overview of different aspects such as amplification method, input DNA amount, yield, application areas, time to results, and compatibility for each version. Additional details can be provided based on specific requirements or applications if needed.

what can be done about the significant delay in delivery?
To address the significant delay in delivery, we recommend staying informed about the shipping status through FedEx tracking. Given that delays are expected during the holiday season, it might be beneficial to consider scheduling shipments after New Year. If a replacement is needed for a specific kit, please provide your shipping address and any relevant catalog or lot numbers to facilitate the process. Our customer support team will keep you updated on when your order is shipped.

why not sell product comparison data
Selling product comparison data could provide valuable insights for customers looking to make informed decisions about which system or vector to choose for their specific needs. However, the focus on direct sales and marketing efforts may not prioritize this type of comparative information at the moment. By concentrating on promoting our own products and systems, the strategy might aim to simplify choices for users rather than complicate them with extensive comparisons. Additionally, resources might be better allocated towards enhancing existing product lines or developing new offerings that align closely with customer requirements in the protein expression field.

know anything specific about binding interactions of magnetic beads with HIV?
The conversation indicates that there isn't specific information available about how HIV binds to magnetic beads. While it is acknowledged that the lentivirus used in their work is based on HIV, the details regarding the binding interactions between the virus and the beads remain unclear. The focus appears to be more on how cells internalize these beads when they are pulled down by a magnet during transduction, rather than on studying the specific interactions of HIV with magnetic beads.

what is the best method for aav purification to maximize yield and what role does media play in this process?
To maximize yield during AAV purification, using a comprehensive approach that includes the AAVpro® Cell & Sup. Purification Kit Maxi (All Serotypes) is recommended. This kit purifies AAV from both lysed cells and supernatant, ensuring you capture as much viral vector as possible compared to methods that only target lysed cells. The choice of media plays a critical role in this process. High-quality cell culture media can enhance cell growth and viability, which directly impacts the amount of AAV produced. Additionally, nutritional components in the media may influence virus production efficiency and particle stability. Therefore, selecting an appropriate media formulation tailored for your specific cell line can significantly improve overall yields during AAV production and subsequent purification steps.

Is this product compatible with UDI kits?
The Unique Dual Index Kits (Cat. Nos. 634752–634756) are designed to work effectively with UDI kits, ensuring compatibility for your needs. If you have any specific requirements or questions about the integration, feel free to ask!

need recommendations for optimal vector systems and cell lines for gram positive strains?
For optimal vector systems and cell lines tailored for gram-positive strains, consider the following recommendations: 1. **Brevibacillus Expression System**: This system is well-suited for expressing proteins in gram-positive bacteria. It offers different vectors that can be used for intracellular or secretory expression depending on your target protein. 2. **Promoter Selection**: Choose a strong promoter suitable for gram-positive organisms to enhance protein expression levels. Look into promoters derived from Bacillus subtilis or other closely related species. 3. **Cleavage Sites**: If using tagged proteins, select vectors with appropriate cleavage sites to facilitate tag removal post-expression, ensuring that the final product is as close to native form as possible. 4. **Cell Lines**: Utilize established strains like Bacillus subtilis or Brevibacillus choshinensis which are often recommended due to their robust growth characteristics and ability to secrete large amounts of recombinant proteins. 5. **Licensing Considerations**: Ensure you review any licensing agreements associated with specific products or systems you plan on using, especially if they involve proprietary technology. Consulting with Technical Support can provide more tailored advice based on your specific applications and experimental design when working with these systems.

can you provide detailed bioinformatic guidance for analyzing UMI portion in the thruplex tag-seq and tag seq flex kits?
Analyzing the UMI (Unique Molecular Identifier) portion in ThruPLEX Tag-Seq and Tag-Seq FLEX kits involves several steps that include data preprocessing, UMI extraction, deduplication, and downstream analysis. Here is a detailed guideline for each of these phases: 1. **Data Preprocessing**: - Ensure your sequencing reads are in a suitable format (e.g., FASTQ). Use tools like `FastQC` to assess the quality of your raw sequencing data. - Trim adapter sequences using tools such as `Trimmomatic` or `Cutadapt`, ensuring you retain parts of the reads containing UMIs. 2. **UMI Extraction**: - Depending on the design of your library kit, identify where the UMI sequence is located within your reads (often at either end). - You can use command-line tools such as `umi-tools` or custom scripts in Python/R to extract UMIs from read sequences efficiently. 3. **Deduplication**: - Utilize tools designed specifically for handling UMIs, like `umi_tools dedup`, which will help you collapse duplicate reads originating from the same molecule based on their UMI tags. - This step improves accuracy by reducing bias introduced during PCR amplification. 4. **Alignment**: - Align deduplicated reads to the reference genome using aligners like `BWA` or `STAR`. Be sure to index with appropriate settings suited for RNA-seq if applicable. 5. **Quantification and Data Analysis**: - After alignment, quantify gene expression levels through counting unique mapped reads per gene using feature counting software such as HTSeq-count or featureCounts. - For differential expression analysis, consider packages in R/Bioconductor such as DESeq2 or edgeR that integrate well with count matrices generated from UMI-based quantification. 6. **Visualization and Interpretation**: - Visualize results using plotting libraries (`ggplot2`, etc.) or dedicated bioinformatics visualization tools (like IGV for viewing aligned reads). - Integrate findings into biological context by comparing against relevant databases or literature regarding expressed genes/proteins. 7. **Quality Control and Reporting**: - Throughout all stages, maintain thorough documentation including parameters used for processing steps so that analyses can be reproduced accurately. These guidelines should provide a comprehensive framework for analyzing UMI portions effectively within ThruPLEX Tag-Seq datasets while ensuring high-quality results throughout your research project.

kit simultaneous dna rna protein purification ffpe?
It seems that there are challenges associated with using a kit for simultaneous DNA, RNA, and protein purification from FFPE samples. The nucleic acids in FFPE tissues tend to be degraded, and the proteins may be denatured, which complicates the extraction process. Based on previous experiences and recommendations, it appears that there isn't an effective kit available specifically designed for this purpose at the moment. If you're looking for reliable results from FFPE samples, you might need to consider separate extraction methods or kits tailored individually for DNA, RNA, or protein isolation instead.

need double order
To assist with a double order, please provide the details of what you would like to reorder. Once I have that information, I can guide you through the process or help ensure your needs are met.

need recommendations for fluorescently labeled primers for microsatellite analysis using capillary electrophoresis?
For microsatellite analysis using capillary electrophoresis with fluorescently labeled primers, it's important to choose a PCR enzyme that ensures high efficiency and specificity. While some enzymes like PrimeStar GXL may not perform optimally with labeled nucleotides, TaKaRa LA Taq® DNA Polymerase is recommended for its ability to amplify long fragments (up to 20kb) and handle high GC content effectively. Additionally, LA Taq offers proofreading activity, providing higher fidelity than wild type Taq. Considering your application focuses on detection rather than requiring the utmost fidelity, prioritizing a high-efficiency polymerase will be essential. If you're concerned about the label placement on the primer (5'-end or middle), ensure that your chosen enzyme can accommodate it well. Overall, utilizing a robust PCR enzyme like LA Taq should enhance your results in microsatellite analysis while supporting the use of fluorescently labeled primers efficiently.

what is the adapter sequence for illumina's universal adapters used in the unique dual index kit?
The adapter sequences for Illumina's Universal Adapters used in the Unique Dual Index Kit can be found in the Illumina Adapter Sequences Document, which provides detailed information on the specific sequences associated with both IDT for Illumina-TruSeq DNA and RNA UD Indexes. For the Unique Dual Indexes (UDIs) U001–U096, these correspond to the standard IDT TruSeq DNA and RNA UD Indexes. The adapters typically include an i5 index sequence of 8 nucleotides and an i7 index sequence of 8 nucleotides as well. To access the exact adapter sequences, please refer to this document: https://support-docs.illumina.com/SHARE/AdapterSequences/Content/SHARE/AdapterSeq/TruSeq/UDIndexes.htm For additional details regarding custom UDIs U097–U384 developed by Takara Bio USA Inc., you may want to consult their user manual available at: https://www.takarabio.com/documents/User%20Manual/Unique%20Dual%20Index%20Kits%20Protocol/Unique%20Dual%20Index%20Kits%20Protocol-At-A-Glance.pdf

What is the difference between the adeno-x adeno rapid titer kit that detects adenovirus particles and the one that detects adenovirus?
The Adeno-X™ Rapid Titer Kit is designed specifically to detect adenovirus particles, particularly focusing on the quantification of viral capsid proteins. This allows for a rapid assessment of viral titer levels in samples. In contrast, kits that detect adenoviruses may identify the presence of the virus itself or its genetic material rather than quantifying just the viral particles. Essentially, one kit measures physical virus components while the other may focus on broader aspects like infection status or genetic verification. Each type serves different purposes depending on what aspect of adenovirus research or diagnostics you are interested in.

how to re-store my soc media stellar competent cells?
To re-store your SOC media stellar competent cells, make sure to keep them sterile throughout the process. You can store the cells at room temperature or in a refrigerator at 4°C. Ensure that they are well-sealed to prevent contamination and maintain their viability. If you have any further questions about handling or storing your competent cells, feel free to ask!

what is the composition of SOC medium provided by the company?
The specific composition of the SOC medium offered by the company is not detailed in the provided information. However, it is noted that any standard SOC formulation should work for your needs. If you require more precise details about the composition, it may be best to contact customer service directly for assistance.

what part number for thruplex tag-seq kit without fragmentation step?
The part number for the ThruPLEX® Tag-Seq FLEX kit without a fragmentation step is R400735. This kit allows for flexible use of unique dual indices (UDIs), which must be purchased separately, and supports input amounts ranging from 250 pg to 200 ng. If you need further assistance or more information, feel free to ask!

what does perfect mean in this context?
In this context, "perfect" likely refers to the ideal way to place an order, either by calling customer support directly or by sending an email. It suggests that these methods are straightforward and effective for obtaining assistance or placing orders with the company.

what conditions should i consider when breaking pellets and how can i avoid issues with vortexing?
When breaking up pellets, it's important to consider a few key conditions to maximize your recovery of AAV particles. Since the AAV-producing cells should be cultured in serum-free medium for optimal purification efficiency, ensure that any residual serum proteins are minimized. To break the pellet effectively without causing damage or inactivation of the virus, you can use gentle methods. Instead of vortexing, which can be risky, try using a pipette tip to carefully break apart the pellet or employ a glass rod or similar tool for manual disruption. This approach allows for more controlled contact with the buffer and helps dissolve as much virus as possible while reducing the risk associated with vigorous mixing. The goal is to facilitate dissolution into solution without compromising viral integrity. By taking these precautions and opting for gentle mechanical disruption rather than vortexing, you can enhance your chances of recovering intact AAV particles from your preparation.

where can I find the capturem maxiprep his-tag purification kit for purchase?
You can find the Capturem His-Tagged Purification Maxiprep Kit available for purchase directly on the Takara Bio website or through authorized distributors. Please note that we do not sell individual components of the kit separately. If you have any further questions about our products, feel free to ask!

hi marianne what to do next?
To proceed, you should first ensure that your murine cells are properly prepared for the ecotropic receptor booster. Make sure to follow any specific protocols related to the treatment or manipulation of these cells. After that, consider running a series of tests to evaluate how effectively the booster is working in enhancing the receptor expression on your cells. If needed, consult relevant literature or colleagues for additional insights or troubleshooting tips based on your observations.

how long should i incubate the retro-x retro concentrator for optimal virus concentration?
For optimal virus concentration using the retro-x retro concentrator, incubation for several days is recommended, especially if you plan to leave it over a weekend. While incubating for just a few hours can be done, it may result in lower performance. The concentrator helps stabilize the virus during storage, and it's been noted that titer levels remain stable even after five days of storage before spinning. However, ensuring proper temperature control is essential; the concentrator works best when stored at 4°C rather than at higher temperatures like 20°C.

is a credit card po required for my order?
A credit card purchase order is not required for your order. You can choose to pay using either a credit card or by submitting a purchase order (PO). If you have any further questions or need assistance, feel free to reach out to customer support.

how much cheaper is it per reaction?
The conversation does not provide specific pricing information for either the SMART-Seq® mRNA HT LP or SMART-Seq® mRNA LP products, so I cannot determine how much cheaper one is per reaction compared to the other. For detailed pricing and cost comparisons, it's best to consult the product catalog or contact customer support directly.

recommendation for larger insert with adeno-x adenoviral system?
For larger inserts using the Adeno-X adenoviral system, it is important to keep in mind that exceeding the recommended insert size can hinder proper packaging into adenoviral particles. The theoretical capacity for a promoterless adenoviral vector allows for an 8 kb total expression cassette, which includes both the promoter and gene. If your insert exceeds this limit, consider optimizing your construct by either reducing its size or splitting it into smaller components that can be packaged separately. Always ensure that you adhere to the guidelines provided in the user manual for best results.

what comparable replacement is available for the longer unavailable components?
The comparable replacement for the longer unavailable components is the In-Fusion® Snap Assembly Starter Bundle (Cat. No. 638945), which includes materials for 10 reactions and is priced at $300.00. This bundle contains: - In-Fusion Snap Assembly Master Mix (Cat. No. 638947) - NucleoSpin Gel and PCR Clean-Up Kit (Cat. No. 740609.10) - Stellar Competent Cells (Cat. No. 636763) - PrimeSTAR Max DNA Polymerase (Cat. No. R045Q) While the Cloning Enhancer (Cat. No. 639615) can be purchased separately for $318, it is not necessary if customers purify their linearized vector and PCR product, as the enhancer's primary function is to facilitate cloning without purification when specific single-band PCR products are used. The new bundle also includes cleanup columns within the NucleoSpin kit, making it a comprehensive option for seamless cloning needs while providing flexibility in handling your PCR products effectively.

what is sensitivity detection
Sensitivity detection refers to the lowest concentration of a substance that can be reliably detected by an assay. In the context of the IL-6 GoStix™ Plus, the sensitivity range is from 3.1 ng/ml to 50 ng/ml, meaning it can accurately detect IL-6 levels as low as 3.1 nanograms per milliliter up to a maximum of 50 nanograms per milliliter. This range indicates how sensitive the test is in identifying varying concentrations of IL-6 in samples.

how can i increase transduction efficiency in neuronal organoids using the virus i'm generating?
To increase transduction efficiency in neuronal organoids using the virus you are generating, consider these strategies: 1. **Use of Lenti-X Transduction Sponge**: The sponge can help bring the lentivirus to the surface of your organoids, enhancing interaction between the virus and target cells. However, it may not assist with penetration into the inner cells of larger organoids. 2. **Optimize Virus Concentration**: Experimenting with different concentrations of your lentivirus can yield better results. Higher titers might improve transduction rates but be cautious about potential toxicity. 3. **Enhance Cell Accessibility**: If possible, modify your organoid preparation or culture conditions to make inner cells more accessible to viral particles. This could involve adjusting size or utilizing techniques that promote dispersion without compromising cell integrity. 4. **Transfection Enhancers**: Consider adding chemical agents or reagents known as transfection enhancers that facilitate viral entry into cells. 5. **Extend Incubation Time**: Prolonging exposure time during infection can give viruses a better chance to penetrate deeper into organoid structures. 6. **Co-culture Systems**: Using co-cultures with helper cell types might enhance overall transduction by providing additional support for viral spread and uptake. 7. **Consult Technical Resources**: Review specific protocols and technical notes related to lentiviral applications on various cell types; gaining insights from others who have successfully used similar methods may also provide useful tips. Reaching out to research and development teams within your organization could offer additional tailored insights based on their experiences with neuronal systems specifically if needed.

possible UMI sequence patterns used for linker disclosure?
The UMI (Unique Molecular Identifier) sequence patterns used in the TCR SMART UMI Oligo consist of 12 random nucleotides, allowing for a theoretical maximum of 16,777,216 unique combinations (4^12). This extensive diversity helps to accurately track individual molecules during sequencing and can aid in distinguishing true biological signals from noise. For specific details on how these UMIs function within your experiments or their application for linker disclosure, please refer to the relevant sections in the SMART-Seq Human TCR User Manual.

what's the best way to concentrate lentivirus for transducing beta cells effectively?
To effectively concentrate lentivirus for transducing beta cells, you can follow these steps: 1. **Ultrafiltration or Precipitation**: Use methods such as ultrafiltration through a 100 kDa filter or polyethylene glycol (PEG) precipitation to concentrate your viral preparation. These techniques help reduce the volume while increasing the titer of your lentivirus. 2. **Centrifugation**: After filtration or precipitation, centrifuge the concentrated virus at high speed to further clear any debris and enhance purity before use. 3. **Determine Optimal Titer**: Perform a small-scale test to determine the optimal multiplicity of infection (MOI) needed for effective transduction without overwhelming your target cells. 4. **Add Slowly and Monitor Cell Health**: When adding the concentrated virus to beta cells, do so slowly and monitor cell health closely since they may be sensitive to high concentrations of viral particles. 5. **Consider Transfection Reagents**: If necessary, consider using transfection reagents that can facilitate uptake if you're dealing with hard-to-transduce hematopoietic cells alongside beta cells. These strategies should help maximize efficiency in achieving successful transduction while minimizing stress on your target cells.

how can I minimize the amount of RNA needed for my reverse transcriptase control while generating a negative control library?
To minimize the amount of RNA needed for your reverse transcriptase control while generating a negative control library, consider the following strategies: 1. **Optimize Reaction Volume**: Reduce the total volume of your reaction, which allows you to use smaller amounts of RNA. Ensure that all reagents are scaled down proportionately to maintain optimal conditions. 2. **Use Concentrated RNA Solutions**: If possible, concentrate your RNA samples before use. This way, you can achieve sufficient reaction concentrations with less total RNA. 3. **Select Specific RNA Targets**: Focus on using only those regions or specific transcripts required for your controls instead of whole transcriptome libraries. 4. **Perform Dilution Series**: Create a dilution series to determine the minimal effective concentration of RNA necessary for successful reverse transcription and amplification. 5. **Employ Alternative Methods**: Consider alternative methods such as using synthetic oligonucleotides or cDNA fragments as controls instead of full-length RNAs if applicable. 6. **Evaluate Enzyme Efficiency**: Use high-efficiency enzymes that require lower input amounts to generate reliable results even at reduced quantities of starting material. 7. **Pooling Samples**: If appropriate, pool multiple small inputs from different sources or replicates rather than relying on larger single inputs for controls. By employing these techniques, you should be able to effectively reduce the quantity of RNA needed while still obtaining valid results in your experiments.

how can we prevent rnase contamination in our dna template prep?
To prevent RNase contamination in your DNA template preparation, it's essential to follow good laboratory practices. Here are some tips: 1. **Use RNase-free reagents and consumables**: Ensure that all buffers, tubes, and pipette tips are certified RNase-free. 2. **Wear gloves**: Always wear gloves when handling samples and reagents to avoid transferring contaminants from your skin. 3. **Clean surfaces**: Regularly clean work surfaces with a solution of 70% ethanol or an appropriate RNase decontamination solution to eliminate any potential residue. 4. **Dedicated equipment**: If possible, use dedicated equipment for RNA work (e.g., pipettes) separate from those used for DNA preparations. 5. **Proper storage**: Store your samples at low temperatures and ensure they are tightly sealed to minimize exposure to the environment where RNases may be present. By adhering strictly to these practices along with following the protocol using appropriate reagents, you can significantly reduce the risk of RNase contamination in your DNA template prep.

what are optimal conditions for transfecting primary human t-cells while maintaining viability and efficiency?
When transfecting primary human T cells while aiming to maintain both viability and efficiency, several key factors need to be considered: 1. **Transfection Reagent**: Utilize the XFect Protein Transfection Reagent, which has been shown to work well for various cell types, including primary T cells. It's important to follow the manufacturer's guidelines regarding optimal reagent-to-protein ratios. 2. **Cell Preparation**: Ensure that T cells are in good health before transfection. You might want to activate them beforehand using anti-CD3/CD28 beads or cytokines to enhance their receptivity. 3. **Media Selection**: Use OPTIMEM media during the transfection process as it supports efficient protein delivery while minimizing toxicity. 4. **Incubation Conditions**: - For adherent T cells, after adding the transfection mixture, remove it after a specified incubation time (typically around 2 hours) and replace with complete medium. - For non-adherent T cells, add two volumes of complete medium directly without removing all liquid from the wells. 5. **Caspase Inhibitors**: Consider pre-treating your T cells with a caspase inhibitor like Q-VD-OPH for about 30 minutes prior to transfection if you anticipate stress responses that could affect cell viability post-transduction. 6. **Post-transfection Care**: After incubating with the transfection mixture, ensure timely washing of the cells before further culturing them in fresh complete medium to reduce any residual cytotoxic effects from reagents used during transduction. 7. **Timing for Assays**: If assessing protein expression shortly after transfection (e.g., within 2 hours), replacing with fresh media should suffice; however, if assays will occur later on, consider gently changing out the medium again at that point. By optimizing these conditions based on specific experimental needs and closely monitoring cell viability throughout the process, successful protein delivery into primary human T cells can be achieved efficiently while maintaining their functionality.

what plasmid system recommend for transfecting difficult cell line with inducible expression?
For transfecting a difficult cell line with inducible expression, the Tet-On 3G lentiviral system would be a strong recommendation. This system allows for controlled expression of your gene of interest and is compatible with challenging cell types like BV2, where lentiviral transduction has proven to be effective. You can choose between options that include fluorescent proteins as well; for example, you might consider using either green or red fluorescent proteins along with your gene. The EF1a promoter is suggested over CMV for this particular application since it may provide better performance in certain cells. The plasmids associated with these systems are available on the manufacturer’s website, allowing you to download maps and relevant documents without an additional purchase necessary after the initial acquisition of the system. This setup not only facilitates working with hard-to-transfect cells but also offers flexibility should you decide to move on to other cell types in the future.

why is gostix not detecting virus?
There could be several reasons why the GoStix is not detecting the virus. One possibility is that the viral titer may be below the detection threshold of approximately 5x10^5 IFU/ml, which means that while there might still be some viral particles present, they are not at a level high enough for detection by the GoStix. Additionally, it's important to ensure that there was a proper flow through the strip; if only a control band appeared without a test band, it indicates that while the assay ran correctly, there may not have been enough target virus in your sample. If feasible, running a positive control on another strip can help confirm whether the test itself is functioning properly. It's also worth noting that certain cell types like HEK cells can easily transduce and might show results even when approaching sensitivity limits. If you're experiencing consistent issues with detection despite these considerations, further investigation into sample preparation or testing conditions may be necessary.

case send new one?
It sounds like there may be an issue with the shipment due to the dry ice situation, and it would be best for your customer to reach out directly to technical support with their order information. This will allow our customer service team to gather the necessary tracking details and work with FedEx on a resolution. If it's determined that a replacement is needed, we can certainly assist further at that point.

what is the shelf life of these lenti products?
The shelf life of the lenti products mentioned is as follows: for product 631276, it is 1 year upon receipt. Product 631232 is also usually stable for about 1 year upon receipt, although some users have reported successful use after its expiration. Lastly, product 631478 has a shelf life of 2 years.

What is the expiration date?
The expiration date is typically included in the Certificate of Analysis. You can check the provided link to view the specific lot number's details, including its expiration date. If you have any further questions or need additional assistance, feel free to ask!

is higher number of cells necessary for better performance using kit?
Using a higher number of cells is not necessary for better performance with the PicoPLEX kits. While it is possible to amplify DNA from more than the recommended number of cells, doing so can compromise the advantages offered by low-input chemistry designed for single-cell genomics. The optimizations made for 1-5 cells have shown promising results, and it's essential to stay within this range to achieve optimal amplification without overwhelming the reaction. Thus, focusing on lower cell numbers aligns better with the kit's intended use and maximizes its effectiveness.

hi mailing address
The mailing address is 2560 Orchard Parkway, San Jose, CA 95131, USA. If you need any further assistance or information, feel free to ask!

kindly provide catalog numbers for phusion high-fidelity pcr master mix and dreamtaq dna polymerase?
The catalog number for the Phusion High-Fidelity PCR Master Mix is typically related to the product from NEB, while the DreamTaq DNA Polymerase's alternative, EmeraldAmp® MAX PCR Master Mix (2X), has a catalog number of RR320A. For further details on these products, you may refer to their respective manufacturer websites.

kindly provide catalog numbers for phusion high-fidelity pcr master mix and dreamtaq dna polymerase?
The catalog number for the Phusion High-Fidelity PCR Master Mix is not explicitly mentioned in the provided conversation, but it can typically be found on the manufacturer's website. For DreamTaq DNA Polymerase, the alternative mentioned is EmeraldAmp® MAX PCR Master Mix (2X), with a catalog number of RR320A. You may want to check the respective product pages for more information and specifics about these products.

who is takarabio?
Takarabio is a biotechnology company that focuses on providing advanced solutions for life sciences research and development. They specialize in areas such as gene synthesis, genetic engineering, and other biotechnological innovations aimed at improving scientific research capabilities. Their services often cater to researchers and organizations looking to advance their work in genetics, molecular biology, and related fields.

Can you help me with tech support for product selection?
Absolutely, I can assist with tech support for product selection. To provide the best recommendations, it would be helpful to know more about your specific needs and preferences. Are you looking for a particular type of product or technology? Additionally, any details regarding budget, intended use, or features that are important to you will help narrow down the options effectively.

hey can you let me know when ian will be available to talk about tbj data kits?
It would be best to reach out directly to Ian or his team for information on his availability to discuss the TBJ data kits. They can provide you with the most accurate and up-to-date schedule.

which genetic analyzer would you recommend for our laboratory needs?
Based on the information provided, it seems that your laboratory is focused on analyzing DNA fragments in the 35-500 nucleotide range. Given this requirement, it's important to choose a genetic analyzer that can efficiently resolve and visualize DNA fragments within this size range. When selecting a genetic analyzer, consider models known for high-resolution capabilities with specific emphasis on smaller fragment sizes. Instruments equipped with advanced detection systems and optimized separation technologies would be beneficial. Ensure compatibility with the dyes you plan to use for labeling, as this could affect visualization results. Additionally, since your ladders have only been tested on agarose gels, an analyzer that is compatible with gel electrophoresis techniques may also be advantageous. Look for analyzers designed for applications involving short DNA fragments and ensure they offer good sensitivity and accuracy in quantifying such lengths. Overall, review options from reputable manufacturers specializing in genetic analysis equipment to find one that meets your laboratory's specific needs effectively.

how can i quantify lentivirus bands without the app?
Quantifying lentivirus bands without the app can be challenging, as the app is designed to facilitate this process. However, you can follow these general steps: 1. **Use a Standard Curve**: If you have access to a standard curve or reference data related to your specific assay, it can help in quantifying your results by comparing the intensity of your test bands against known values. 2. **Visual Comparison**: You might visually compare the Control (C) line and Test (T) line on your test strip. The C line should always be darker than the T line for valid results. The relative darkness of these lines can give an approximate indication of viral concentration. 3. **Image Analysis Software**: Consider using image analysis software that allows for densitometric analysis of gel images or strips if applicable. This software often provides quantitative measurements based on pixel intensity. 4. **Consult Documentation**: Refer to any available documentation or product FAQs from manufacturers that may provide guidance on manual quantification methods. 5. **Experimental Replicates**: Conduct multiple tests and analyze replicate results statistically for more reliable data interpretation. For additional resources, check out relevant sections in scientific literature or manufacturer guidelines that discuss alternative methodologies for determining viral concentrations without relying solely on mobile applications.

Can you provide the specific catalog numbers and a quote for the packaging of AAV with the required plasmids?
For the packaging of AAV2, here are the specific catalog numbers and their respective prices: 1. **AAVpro® Packaging Plasmid (AAV2)** - Catalog Number: 6234, Price: USD $1055.00 2. **VP/AAP** - Same catalog number as above: 6234. 3. **AAVpro® Purification Kit Maxi (All Serotypes)** - Catalog Number: 6666, Price: USD $821.00. 4. **AAVpro® Titration Kit (for Real Time PCR) Ver.2** - Catalog Number: 6233. Additionally, consider using the **AAVpro® 293T Cell Line**, which has higher transfectibility for better viral titers; details can be found on the manufacturer's website. If you have any further questions or need additional assistance with your order, feel free to reach out for support later on.

is lentix included in the discount code for my order?
To determine if Lentix is included in the discount code for your order, please check the terms and conditions associated with the code or contact customer support directly for clarification. If you have any specific details about the promotion, feel free to share them as it may help us provide a more accurate answer.

What are the possible gDNA starting templates for human TCR sequencing kits?
For human TCR sequencing kits, possible genomic DNA (gDNA) starting templates typically include: 1. **Peripheral Blood Lymphocytes (PBLs)**: Isolated from whole blood samples, PBLs provide a rich source of T cells for profiling. 2. **T Cell Enriched Samples**: These can be obtained through methods such as magnetic bead separation or flow cytometry to isolate specific T cell populations. 3. **Tissue Biopsies**: gDNA extracted from solid tumors or lymphoid tissues can also serve as templates for TCR sequencing. 4. **Cell Lines**: Established human T cell lines may be used when studying specific characteristics of known clones. 5. **Primary Tumor Samples**: In oncology studies, tumor-infiltrating lymphocytes (TILs) can be analyzed for their TCR repertoire. It’s important to ensure that the quality and integrity of the gDNA are suitable for downstream applications in order to obtain reliable results in TCR profiling.

can i buy tetr monoclonal antibody conjugated fitc and the unconjugated version separately?
It appears that the tetr monoclonal antibody is available in two sizes: purified and suspended in PBS. However, it seems that there is no option for a conjugated version, such as FITC, available for purchase. If you are specifically looking for an unconjugated form, you can definitely obtain that. Additionally, the antibody should be compatible with various conjugation chemistries if you wish to perform your own conjugation later on. For more details about product specifications and availability, checking the provided links may be helpful.

where can i buy the dna analysis sell kit?
You can purchase DNA analysis cell kits from various scientific supply companies and online marketplaces that specialize in laboratory reagents and assay kits. It’s a good idea to check the websites of reputable manufacturers or distributors in your region for availability. Additionally, you may find them through academic suppliers or biotech retailers.

what is concentration product?
The concentration of a product, particularly in the context of antibodies, refers to the amount of antibody present per unit volume. In your case, since the Cas9 antibody is not purified and lacks a specific concentration value, you will rely on recommended working dilutions for use in experiments like Western blotting or immunocytochemistry. For instance, with the provided titer range for Western (1:1000-1:5000), this indicates how much you should dilute the serum containing the antibody before application in your assays. Typically, more concentrated solutions yield stronger signals but may also lead to non-specific binding if used excessively; therefore, following suggested dilution recommendations helps optimize results while minimizing background noise.

wanted know columns compatible transfection grade nucleospin kit?
The NucleoSpin® Plasmid Transfection-grade columns are specifically designed for that kit and are not available for separate purchase. Unfortunately, Macherey Nagel does not recommend using other columns as replacements. It's also worth noting that they typically provide additional buffers to support the transfection process. If you have further questions about compatibility or need assistance with your experiments, feel free to ask!

how can I troubleshoot issues with overlapping primers in site-directed mutagenesis?
To troubleshoot issues with overlapping primers in site-directed mutagenesis, start by verifying the design of your primers using a reliable primer design tool. Check for potential problems like primer dimer formation or hairpin structures that could interfere with amplification. Ensure that the overlap between primers is approximately 15 bp, as this length generally facilitates successful In-Fusion reactions without complications. Additionally, confirm that the overlapping sequences are positioned at the 5' ends of the primers to minimize any risk of forming primer dimers during PCR. Conducting your In-Fusion reaction at around 50°C can also help improve specificity and efficiency. If issues persist, consider adjusting annealing temperatures or redesigning problematic primers while keeping these factors in mind.

what are the recommended mammalian expression vectors for transient expression in a one or two plasmid strategy for my synthetic igg fusion protein?
For transient expression of your synthetic IgG fusion protein, you might consider using either bidirectional promoter vectors or bicistronic IRES vectors to achieve optimal expression of both heavy (H) and light (L) chains. Bidirectional promoter vectors allow for the simultaneous expression of two coding sequences from a single plasmid; however, they may not express the two proteins equally. In this case, using two separate plasmids could provide more balanced expression levels for the H and L chains. Bicistronic IRES vectors can also be an effective option since they enable co-expression of multiple genes by including an Internal Ribosome Entry Site (IRES), which facilitates translation of downstream coding sequences. Reviewing the following links may help you choose the best approach: - Bidirectional Promoter Vectors: [Link](https://www.takarabio.com/products/gene-function/mammalian-expression-plasmids/bidirectional-promoter-vectors) - Bicistronic IRES Vectors: [Link](https://www.takarabio.com/products/gene-function/mammalian-expression-plasmids/bicistronic-ires-vectors) - pIRES Vector Information: [Link](https://www.takarabio.com/documents/Vector%20Documents/pIRES%20Vector%20Information.pdf) Choosing between these strategies will depend on your specific needs regarding protein yield and balance in expressing both chains effectively.

aav extraction sol'n confirm mechanisms kit aav extract aav supernatant culture's medium total virus suspension final confirmation proceed
The AAV extraction process involves several key steps to ensure effective isolation of the virus from the culture medium. Initially, you'll want to concentrate your viral particles using the Amicon Ultra-4 filter with a 100 kDa cut-off, which retains larger molecules while allowing smaller contaminants to pass through. After filtering, it's important to wash the membrane by adding Suspension Buffer (which acts as a wash buffer) and spinning it down again. This helps resuspend any remaining virus that may be stuck on top of the membrane while discarding unwanted filtrate. The majority of AAV will remain on top due to its size. Following this, you would add Cryonase for one hour, which digests any genomic DNA outside of the AAV particles in order to minimize contamination risks from host DNA. Finally, Precipitator A/B is introduced to precipitate your concentrated AAV virus from solution. Once these steps are completed and you've confirmed no significant contamination remains, you can transfer your final viral suspension into a new tube for downstream applications or further analysis. If there are specific details or additional questions regarding this process you'd like assistance with, feel free to ask!

Can you ship my order on Monday instead?
Your order is likely to ship on Monday for overnight delivery, as we do not process shipments over the weekend or on holidays. If you need your order to be shipped specifically on Monday, that should align with our shipping schedule.

how do mn kits compare to qiagen allprep dna rna kits for my applications?
When comparing the NucleoSpin® TriPrep kits to Qiagen's AllPrep DNA/RNA kits, both options offer effective isolation of nucleic acids. The NucleoSpin TriPrep allows for simultaneous extraction of DNA, RNA, and protein from various starting materials—such as cultured cells and tissues—making it versatile for applications that require all three macromolecules. If your application does not require protein isolation, you can simply skip that step. On the other hand, Qiagen’s AllPrep kits are designed to provide a streamlined process for isolating both DNA and RNA from the same sample but may not include protein purification in their standard workflow. The NucleoSpin RNA/DNA Buffer Set is an economical choice if you're looking to isolate just DNA and RNA from specific tissue types while providing more flexibility by allowing combinations with various NucleoSpin RNA products. Ultimately, your choice between these two will depend on specific needs such as whether you need protein extraction or prefer separate processing steps for different nucleic acids. Consider factors like cost-effectiveness, ease of use, and compatibility with your downstream applications when making your decision.

what amount ng total rna should I use to start cDNA synthesis for qPCR?
For cDNA synthesis in preparation for qPCR, you can start with a range of 5 ng to 5 μg of total RNA. The specific amount may depend on the abundance level of your target mRNA transcript. If the mRNA is well-expressed in your tissue sample, starting with as little as 5 ng could be sufficient. After synthesizing cDNA, it's advisable to test several dilutions when preparing for qPCR to accurately determine the expression level of your cDNA.

where is my order?
To check the status of your order, it's best to contact customer service directly for assistance. You can call them or send an email to inquire about your order's whereabouts. They will be able to provide you with the most accurate and timely information regarding your shipment.

provide kit assay it's atp measurement assay?
Unfortunately, we do not offer a kit specifically for ATP measurement. However, I can reach out to Cellartis to see if they have more information regarding the ATP assay kit they used. If you're interested in that option or need assistance with something else related to assays, please let me know!

what should i do if the pcr reagents are frozen for longer than overnight?
If your PCR reagents have been frozen for longer than overnight, it's best to thaw them gently at 4°C or on ice. After thawing, mix the reagents thoroughly by pipetting up and down or vortexing briefly to ensure they are well combined. Check the expiration dates and inspect for any signs of degradation before use. If everything looks good, you can proceed with your PCR reactions as planned. Always remember to minimize freeze-thaw cycles in the future to maintain reagent integrity.

How can I reset my login password to place an order?
To reset your login password, you can typically find a "Forgot Password" link on the login page of the Takara Bio website. Clicking this link will guide you through the process to reset your password. If you're unable to log in or need immediate assistance with placing an order, consider calling customer support at the provided number during their business hours for help.

what should we do if there is no pellet formation during centrifugation?
If there is no pellet formation during centrifugation, consider the following steps: 1. **Check Your Medium**: If you are using serum-free medium, the pellet may be less visible or even "transparent." You might want to try adding some serum to enhance precipitation. 2. **Increase Serum Concentration**: Adding more serum can help form a larger pellet. Starting with 5% and potentially increasing up to 10% could improve recovery. 3. **Centrifugation Parameters**: Ensure that you are using appropriate spin speed and time for your centrifugation process. It’s also important that the precipitation occurs overnight at a low temperature (4 degrees Celsius). 4. **Visual Marking**: Carefully mark out the area of where the pellet should form in your conical tube before centrifugation. This will aid in locating it after spinning. 5. **Supernatant Handling**: If you still cannot see any signs of a pellet, pour the supernatant into a new conical tube to check for any residual virus particles. 6. **Rinsing Technique**: Consider rinsing out the original conical tube with an appropriate buffer solution to recover any potential dissolved virus from previous samples. 7. **Confirm Virus Presence**: Conduct titer assays on both your precipitated material and supernatants to confirm if virus recovery has occurred despite not visually spotting a pellet. By implementing these strategies, you may increase your chances of recovering viral particles effectively even when initial visual cues suggest otherwise.

why is the sequencing data not aligning with the combined tcra tcrb results?
There could be several reasons why the sequencing data is not aligning with the combined TCR alpha (TCRA) and TCR beta (TCRB) results. One potential issue might be related to the quality of the samples or PCR amplification, as variations in starting material can lead to inconsistent results. Additionally, if there are differences in primer specificity or inefficiencies during amplification, this could affect alignment. Another factor to consider is the bioinformatics analysis itself; misalignment may occur due to errors in sequence processing or filtering steps within the pipeline. It’s also essential to ensure that any reference sequences used for alignment are appropriate and accurately represent the target regions. Lastly, it might help to review whether there have been any updates or changes made in protocols since your last analysis that could influence results. Utilizing tools like Cogent Immune Profiling software may provide further insights into discrepancies by offering advanced analytical capabilities for immune profiling data.

can you suggest a more cost-effective option for long repetitive transposons using la taq?
For amplifying long repetitive transposons, while LA Taq is a good choice for challenging templates due to its high fidelity and ability to handle difficult sequences, you might consider using a more cost-effective polymerase that has been reported to perform well with similar applications. GXL Premix could be an alternative worth exploring, as it offers robust amplification capabilities in challenging conditions. Additionally, based on the findings from the reference you provided, it's noted that PrimeSTAR and AccuPrime Pfx showed better performance with higher concentrations of correctly sized fragments compared to other polymerases. If budget allows, testing these options alongside your current setup could help identify the best balance between cost and performance for your specific needs.

which system is better for inducing protein expression in primary neuronal cultures, proteotuner or tet-one?
The choice between the proteotuner system and the TetONE inducible system for inducing protein expression in primary neuronal cultures depends on several factors, including the specific requirements of your experiment. The proteotuner utilizes a CMV promoter to express destabilized proteins of interest (POIs), which is functional in primary neurons but may not be optimal. In contrast, the TetONE system employs an hPGK promoter to drive expression of the tet activator protein, allowing for activation through a Tet response element (TRE3G promoter). This combination can provide more controlled and potentially stronger induction compared to other systems. Research indicates that different promoters have varying efficiencies when transducing central nervous system cells, so it may be beneficial to consider existing literature and perform preliminary tests with both systems. Ultimately, if precise control over timing and levels of expression is critical for your study, the TetONE system might offer advantages. However, if you require rapid degradation of induced proteins or simpler setups without stringent regulation needs, then the proteotuner could suffice. Evaluating both options based on experimental goals will lead you to make a better-informed decision.

wondering data comparing cell lysis methods and brands similar to sonication
When comparing cell lysis methods and brands similar to sonication, it's important to consider several factors such as efficiency, ease of use, and the type of cells being lysed. Sonication is a widely used technique that employs high-frequency sound waves to disrupt cell membranes, resulting in effective lysis for various cell types. Alternatives to sonication include mechanical methods like bead beating or homogenization, which can also achieve efficient cell disruption but may require different equipment and optimization for specific applications. Chemical lysis buffers like xTractor Buffer are designed specifically for protein extraction and can be more gentle on proteins than physical methods. Each method has its advantages: sonication provides rapid results with good reproducibility but may generate heat leading to protein degradation if not carefully controlled. On the other hand, chemical lysis buffers might be milder but could take longer depending on the protocol. It’s beneficial to evaluate specific data from studies or product comparisons related to yield and purity when choosing a method or brand for your needs. Additionally, user experiences shared through scientific literature or forums can provide insights into how these methods perform under different conditions.

what else can i do to troubleshoot this issue?
To troubleshoot the issue further, consider the following steps: 1. **Check Reagents**: Ensure that all reagents are fresh and properly stored. Look for any signs of degradation or contamination. 2. **Optimize Primer Design**: If you have a different set of primers known to work, try using those with your current template to see if amplification occurs. 3. **Run Controls**: Include both positive and negative controls in your experiments. The positive control should yield a clear result, while the negative control should confirm no contamination is present. 4. **Adjust Cycling Conditions**: Review and possibly optimize your PCR cycling conditions (annealing temperatures, extension times) based on the specific requirements of your primer-template combination. 5. **Gel Analysis**: If you haven't already, run a gel on any products obtained from previous reactions to check for expected band sizes or non-specific amplifications. 6. **Consult Colleagues**: Don’t hesitate to ask colleagues for their insights; sometimes another perspective can help identify overlooked issues. 7. **Review Equipment Calibration**: Ensure that the equipment used (like thermocyclers) is calibrated correctly and functioning as intended. 8. **Consider Template Quality/Concentration**: Make sure that you're using high-quality DNA templates at an appropriate concentration—too much or too little can inhibit amplification. These steps should help narrow down potential problems in your reaction setup!

do you have any recommendations for non-mouse secondary antibodies that are still available?
There are several options for non-mouse secondary antibodies that you might consider. Depending on your specific needs, you could look into rabbit or goat-derived secondary antibodies, which are commonly used and widely available. Additionally, some companies offer polyclonal or monoclonal antibodies that target various primary antibody species. It may also be helpful to check with different suppliers for their current inventory of secondary antibodies suitable for your applications in tissue sections or western blotting. If you're looking for something specific, reaching out to the product manager might provide more clarity on availability and alternatives.

long expect shipping process
The shipping process typically depends on various factors, including the time of order processing and any potential disruptions like holidays or weather conditions. In this case, it seems that after the request is processed, the replacement would normally ship out the same day for next-day delivery. However, due to current circumstances such as a holiday week and possible weather issues, there may be some delays. It's best to keep an eye on updates from customer service for more accurate information regarding your specific order.

am i missing essential step in the extraction process affecting yield?
It sounds like you have a solid understanding of the extraction process, but there are several critical steps that can indeed affect yield. Here are a few points to consider: 1. **Sample Preparation**: Ensuring that cells are completely pelleted and the culture medium is fully removed is essential, as residual medium can introduce contaminants that may impact yield or purity. 2. **Centrifugation Parameters**: The rpm used during centrifugation is crucial; while 400 g for 10 minutes might be acceptable, make sure it's suitable for your specific cell type to avoid damaging the cells. 3. **Flash Freezing**: Flash freezing in liquid nitrogen immediately after harvest helps preserve RNA integrity and minimizes degradation or loss of material. Ensure samples do not thaw before contact with lysis buffer, as this could compromise yield. 4. **Homogenization**: Homogenizing frozen samples rather than thawed ones can prevent RNA degradation and improve overall extraction efficiency. 5. **Lysis Buffer Addition Timing**: Adding lysis buffer before allowing any thawing ensures that cellular contents are effectively released without degradation from environmental exposure. 6. **Disruption Methods**: Consider reviewing any disruption methods you're using (e.g., mechanical homogenization versus enzymatic) as they can significantly influence both yield and quality based on how well they disrupt cell membranes without causing excessive shearing of nucleic acids. By carefully evaluating each step in your protocol—particularly those related to sample handling post-harvest—you should be able to enhance both the yield and purity of your extracted material.

am i missing essential step in protocol that's affecting my RNA extraction yield?
Based on the details you've shared, there are a few critical steps to ensure optimal RNA extraction yield. Here are some essential considerations: 1. **Sample Handling**: It’s important to flash freeze samples in liquid nitrogen immediately after harvesting to prevent degradation of RNA. Ensure that your samples do not thaw before they come into contact with the lysis buffer. 2. **Centrifugation**: When preparing your cell pellet, it's crucial to completely remove the culture medium as it contains proteins that can interfere with downstream applications. A centrifugation speed of 400 g for 10 minutes is generally acceptable, but always confirm that cells remain intact and properly pelleted. 3. **Lysis Buffer Addition**: If you're working with lysed cells or a cell pellet, adding the lysis buffer directly after flash freezing (or while still frozen) can enhance yield and purity by quickly disrupting cellular components without allowing RNA degradation. 4. **Storage Conditions**: Post-extraction storage should be at -70 °C if you plan to store samples for more than short-term use; this helps maintain sample integrity over time. 5. **Homogenization**: Homogenizing in a frozen state can assist in achieving better yields since it prevents any potential loss due to thawing prior to lysis. If you follow these guidelines closely and ensure each step is executed carefully, it should help improve your RNA extraction yield significantly. Consider reviewing each step of your protocol against these points to identify any areas for improvement.

What alternatives are available for formaldehyde fixation?
There are several alternatives to formaldehyde fixation that can be considered, depending on the specific requirements of your study. Some commonly used alternatives include: 1. **Ethanol Fixation**: Ethanol is often used for preserving nucleic acids and proteins in samples. It provides good preservation while minimizing crosslinking. 2. **Methanol Fixation**: Similar to ethanol, methanol can preserve cellular structures while causing less protein crosslinking compared to formaldehyde. 3. **Acetone Fixation**: Acetone is effective for preserving certain antigens and may be suitable for immunohistochemical applications. 4. **Bouin's Solution**: This fixative contains saturated aqueous picric acid, formaldehyde, and acetic acid; it's particularly good for soft tissues but does contain formaldehyde. 5. **Zinc Formalin**: Although it still involves formaldehyde, zinc salts can enhance tissue morphology and antigenicity compared to traditional buffered formalin alone. 6. **Paraffin Embedding Methods without Crosslinkers**: Techniques that avoid harsh chemicals or crosslinkers during embedding can help maintain DNA quality for downstream analysis. Choosing the appropriate fixation method will depend on factors such as the type of sample being processed and the intended downstream applications (e.g., histology vs molecular analysis). Always consult relevant protocols or literature specific to your research needs when selecting a fixation method.

what final formulation should I use for the concentrated purified lentiviral vectors intended for mouse studies?
For concentrated purified lentiviral vectors intended for mouse studies, it is advisable to use a buffer exchange method with a 100 kDa MW cutoff Amicon or similar filter unit. This will help remove residual concentrator components and co-concentrated serum proteins that may interfere with your experiments. After concentrating the viral particles, consider using an appropriate buffer such as PBS (phosphate-buffered saline) or another suitable formulation that supports viral stability and efficacy in vivo. It's important to ensure that any remaining contaminants from the culture media are minimized before administration into mice. If you have specific requirements based on your experimental design, adjusting the final formulation accordingly would be beneficial.

what info do you have on hepatocyte maintenance?
The conversation indicates that there is customer feedback confirming the effectiveness of a product with hepatocytes from monkey, rat, and dog species. Additionally, it seems like there might be more recent information available or confirmation on this topic through further inquiries to team members. If you're looking for specific details on hepatocyte maintenance techniques or protocols, it may be beneficial to reach out to colleagues who specialize in this area or check updated resources within your organization.

let know whether shorter overlap solves problem
Shorter overlaps, ideally around 21 bases or fewer, can enhance the efficiency of your cloning process by minimizing potential mispriming and increasing the likelihood of successful amplification. Since longer overlaps may not provide significant advantages and could complicate the reaction, reducing the overlap to a shorter length might help address any issues you’re experiencing. It's always good to experiment with different conditions, including checking for high GC regions in your gBlock that might be causing smearing. Using high-fidelity polymerases like PrimeStar Max is also advisable to reduce errors during amplification.

what's the tracking number for the new infusion kit?
The tracking number for the new In-Fusion cloning kit will be provided by our customer service team once the package is shipped. They should send you an email with all the details, including the tracking information from FedEx. If you have any other questions or need further assistance, feel free to ask!

need help troubleshooting cloning with added nucleotides and vector overlaps?
To troubleshoot your cloning with added nucleotides and vector overlaps, it's important to ensure that the inserts have the correct overlap with the vector. Since you need a 20 base pair (bp) overlap at each end of your insert, you should confirm that you've included an additional 40 bp in total—20 bp on each side—to facilitate this overlap. For linearizing your vector, make sure you're using appropriate restriction enzymes or methods compatible with your cloning strategy. This step is crucial for allowing proper ligation or In-Fusion reaction. Given the mixing instructions, you'll want to maintain a 2:1 molar ratio of insert to vector when preparing your In-Fusion reaction. For example, if you're mixing 110 ng of vector along with two inserts (one being 1800 bp at 48 ng and another being 1600 bp at 42 ng), double-check that these amounts align correctly based on their sizes and concentrations. Lastly, if you're looking for precise primer designs that include those necessary overlapping bases for both ends of your inserts, it’s recommended to utilize a Primer Design Tool available through Takara's website. This will help generate specific primers needed for amplifying your inserts with the required overlaps before proceeding further in the cloning process.

need help figuring out how to properly design overlap and cloning strategy for my vector and inserts?
To design the overlap for your cloning strategy, it's important to ensure that your inserts have a 20-base overlap with the vector. Since you mentioned using gBlocks for your inserts, you should add an additional 20 bases at each end of both inserts (1800 bp and 1600 bp) to create the necessary overlaps. For linearizing the vector, typically this can be done through restriction digestion or PCR amplification if you're using a specific method designed for your application. Be sure to confirm that it is appropriately linearized before proceeding. Regarding mixing quantities, you will want to follow the recommended molar ratio of insert to vector. In this case, you'll mix 110 ng of linearized vector with 48 ng of one insert (1800 bp) and 42 ng of another insert (1600 bp). It’s crucial to calculate these amounts based on their respective lengths so that they are in a proper molar ratio. So in summary, by adding those extra bases at each end of your inserts for the required overlaps and ensuring correct quantities are used according to their sizes during mixing, you’ll set up a solid foundation for successful cloning.

what could be causing the dna contamination in my rna extraction?
DNA contamination in RNA extraction can result from several factors. One common cause is the presence of excess nuclei, especially if a higher number of cells were loaded onto the column than recommended (up to 10^5 cells). Additionally, insufficient or ineffective DNase treatment can lead to incomplete digestion of genomic DNA. Factors such as using an inactive DNase due to improper storage or repeated use may also contribute. The conditions during DNase treatment are crucial; for optimal activity, it should be performed at room temperature (~20°C) rather than at lower temperatures that could diminish its effectiveness. Moreover, ensuring that the DNase reaction mixture is applied directly to the center of the membrane and allowing adequate incubation time will enhance its efficiency. If you're measuring DNA contamination through sensitive methods like PCR, consider alternative approaches or performing additional rounds of DNase treatment. It's also beneficial to explore variations in your protocol—like adjusting temperatures during treatments—to see if this affects RNA quality while addressing contamination issues.

wondering if the isolation kit will effectively separate plasmid dna from gDNA in my transfected cells?
The effectiveness of an isolation kit in separating plasmid DNA from genomic DNA (gDNA) in transfected cells largely depends on the specific kit's design and protocol. Many commercial kits are optimized to purify plasmid DNA while minimizing contamination with gDNA. However, if you're isolating DNA from transfected cells after a few days, there may be some residual plasmid contamination due to the transient nature of plasmids; they often coexist alongside cellular nucleic acids before being degraded. In general, standard protocols for isolating plasmid DNA can help achieve good separation, but it's advisable to check if the kit specifically mentions its ability to differentiate between these two types of genetic material. Additionally, consider factors such as transfection efficiency and timing since these can influence your results. If you have further concerns or need specific recommendations based on your experimental setup, consulting product literature or technical support could provide additional guidance tailored to your needs.

ISO certificate
https://www.takarabio.com/documents/Corporate/TBUSA%20ISO%2013485-2016_Final%20Cert.pdf

Takara Bio Address HQ
Takara Bio USA, Inc. 2560 Orchard Parkway, San Jose, CA 95131 United States

what recent publications using adenoviral system can help convince reviewers for my grant application?
To find recent publications that utilize adenoviral systems, particularly those incorporating the Tet-On 3G system, it would be beneficial to explore scientific journals and databases such as PubMed or Google Scholar. Look for reviews or research articles published in the last few years that highlight advancements or applications of adenoviral vectors in gene therapy, cancer research, or other relevant fields. Additionally, you can refer to the citation/application lists provided by companies specializing in these technologies. For example, resources from providers like Takara Bio may include case studies or application notes demonstrating successful uses of their adenoviral vector systems. It's also useful to search for specific keywords related to your project focus combined with terms like "Tet-On," "adenoviral vector," and "gene expression." This strategy will help you identify cutting-edge studies that could strengthen your grant application by showcasing current trends and successes within the field.

why isnt the email coming for my account login reset?
There could be several reasons why you're not receiving the email for your account login reset. It's possible that the email is getting caught in your spam or junk folder, so it's worth checking there. Additionally, ensure that you entered the correct email address associated with your account when requesting the reset. If you've done both and still haven't received it, consider reaching out to customer support directly for further assistance. They can help investigate any issues related to your account or the reset process.

What do I need to start a laboratory?
To start a laboratory, you will need to consider several key components: 1. **Define Your Focus**: Determine the specific area of research or experimentation your lab will specialize in. This could range from biotechnology to environmental science. 2. **Funding**: Research grants are a primary source of funding for academic labs. Identify potential grant opportunities and understand the application process. 3. **Location and Space**: Secure an appropriate space that meets safety regulations and is equipped for your intended experiments, whether it’s a dedicated lab facility or a portion of existing space. 4. **Equipment and Supplies**: Based on your research focus, procure essential equipment such as microscopes, centrifuges, incubators, and any specialized tools necessary for your work. 5. **Compliance and Safety Protocols**: Familiarize yourself with local regulations regarding laboratory operations including biosafety protocols if you're working with biological materials. 6. **Staffing**: Depending on the scale of your lab, consider hiring skilled personnel like researchers or technicians who can assist in conducting experiments. 7. **Networking**: Build relationships within the academic community to gain insights into best practices and collaborate on projects which could enhance resources available to you. By carefully addressing these areas, you'll be well-prepared to establish a successful laboratory tailored to your research interests.

is sample available for kit nucleospin triprep?
You can request a free sample of the NucleoSpin Triprep kit online or by contacting your territory manager. For more details, you can visit the company's special offers page.

recent publications using adenoviral system for grant application?
For recent publications using adenoviral systems, it's recommended to explore the citation/application list provided by Tet Systems, as they maintain a comprehensive database of studies utilizing their technology. You may also conduct literature searches in scientific databases like PubMed or Google Scholar with keywords such as "adenoviral vector," "Tet-on system," and specific applications relevant to your grant proposal. Additionally, reviewing journals focused on gene therapy and molecular biology can yield valuable insights into current research trends and findings related to adenoviral vectors. If you're looking for specific examples or further information, I would be happy to assist in identifying key papers or resources that align with your interests.

how can i place an order?
To place an order, you can reach out directly through the company's designated contact methods. You may call their customer service number and choose the appropriate option for orders, or you can send an email to inquire about placing your order. This will ensure that you receive prompt assistance and guidance on the ordering process.

How do lenti-x and lenti gostix work in detecting retroviruses like lentivirus?
Lenti-X and Lenti-Gostix are tools designed for the detection and study of lentiviruses, particularly HIV-1. Lenti-X systems typically utilize a reporter gene that can be expressed in host cells infected by the virus, allowing researchers to monitor viral infection and replication through measurable signals, such as fluorescence or luminescence. On the other hand, Lenti-Gostix may refer to specific kits or assays that provide methodologies for detecting lentiviral particles or assessing transduction efficiency in various cell types. These products often rely on similar principles of using selectable markers or detectable reporters linked to viral components. While these tools are effective for studying HIV-1 related lentiviruses, they do not currently have retrovirus-specific versions available. Ongoing research efforts may lead to developments aimed at expanding their application to other retroviruses in the future.

what transduction methods have you found to be most effective for immortalized kidney cell lines and primary human kidney cells?
It appears that clonal kidney cell lines, such as HEK cells, work very well with the transduction methods you've been using. However, primary human kidney cells can be more challenging due to their sensitivity and lower transduction efficiency. The sponge method you mentioned seems to be effective across various cell types tested, which is promising for both immortalized and primary cells. It might be worth exploring different optimization techniques or vectors specifically designed for primary cells if you're looking to improve outcomes further.

what differences umi udi and how do they impact rna sequencing?
UMIs (Unique Molecular Identifiers) and UDIs (Unique Dual Indexes) serve distinct roles in RNA sequencing, each contributing to data quality and analysis. UMIs are short sequences incorporated into cDNA during the first-strand synthesis process. Their primary function is to help distinguish between unique molecules, allowing researchers to identify and eliminate PCR duplicates and sequencing errors. This improves the accuracy of transcript quantification, particularly in low-input RNA samples or when analyzing complex transcripts. By collapsing reads with identical UMIs during analysis, one can achieve a clearer picture of gene expression levels. On the other hand, UDIs are used for demultiplexing sequencing libraries that contain multiple samples within a single run. Unique Dual Indexing allows for precise identification of each sample based on its specific index combinations during sequencing. This ensures that reads from different samples do not get mixed up, facilitating accurate downstream analysis. In summary, while UMIs enhance the reliability of transcript counts by addressing duplicates and errors at the molecular level, UDIs ensure correct sample identification during multiplexed sequencing runs. Together they significantly improve both the quality and resolution of RNA-seq data analyses.

Can you help me find the email for the sales department to restore previous quotes and place an order?
It sounds like you're looking to reach out to the sales department for assistance with restoring previous quotes and placing an order. It might be best to check the company’s official website or contact their general support line for the appropriate email address. If you have any specific contacts in mind, following up directly with them could also help expedite your request.

request free sample xfect mes cells?
It seems that the Sales team is currently unavailable to assist with your request for a free sample of Xfect MEF cells. However, I can forward your inquiry to them, and they will reach out to you via email as soon as possible. If there's anything else you need in the meantime, feel free to let me know!

what could be the reason my fusion cloning kit is not giving colonies in test samples even though the positive control works?
There could be several reasons why your fusion cloning kit is not yielding colonies in the test samples, even if the positive control is working. Here are a few possibilities to consider: 1. **DNA Quality and Quantity**: Ensure that the DNA you're using for the test samples is of high quality and within the optimal concentration range recommended by the protocol. 2. **In-Fusion Reaction Conditions**: Review your reaction setup to confirm that all components were added correctly and in appropriate amounts. Any deviation from established protocols can impact results. 3. **Transformation Efficiency**: Check whether you are using competent cells with high transformation efficiency, as lower efficiency could lead to fewer colonies on plates. 4. **Recovery Medium**: Confirm that you are using SOC medium or another suitable recovery medium after heat shock for cell recovery; this step is crucial for maximizing colony formation. 5. **Cloning Strategy Issues**: If there are any issues with the insert or vector compatibility, such as incorrect restriction sites or sequences, it may prevent successful ligation and transformation. 6. **Environmental Factors**: Consider factors like temperature fluctuations during storage or handling that might affect component stability or activity. Investigating these areas may help identify potential issues affecting your test samples' performance in comparison to the positive control.

why bioanalyzer preferred customer tapestation?
The Agilent 2100 Bioanalyzer is preferred over the TapeStation for several reasons. One key advantage is that the Bioanalyzer provides a distinct cDNA peak, which facilitates clearer interpretation of size distribution and yield during analysis. This clarity can alleviate some concerns customers or core facilities may have regarding the expected broad size distribution associated with SMART-Seq libraries. Additionally, while both systems are capable of analyzing samples, the Bioanalyzer's High Sensitivity DNA kit offers reliable results that many users find more straightforward to interpret compared to those from the TapeStation. The TapeStation’s screen tape profiles may be subject to varying interpretations, making it less desirable in situations where precise understanding of library quality and characteristics is crucial. Overall, for applications requiring detailed assessment of cDNA and library preparations—especially when working with single-cell RNA sequencing—the consistency and reliability offered by the Bioanalyzer often make it a preferred choice among researchers.

is the reconstituted standard stable for long-term storage?
The reconstituted p35 stock, at a concentration of 400 ng/ml, is stable for up to 4 weeks when stored at 4˚C. For long-term storage beyond this period, it would be advisable to consider alternative preservation methods or conditions, such as freezing the stock.

which polymerase would you recommend for high fidelity amplification of difficult sequences?
For high-fidelity amplification of difficult sequences, Takara Ex Premier DNA Polymerase is a strong recommendation. It has shown consistent performance across various genomes and is also noted for its inhibitor tolerance, making it suitable for challenging samples. In studies comparing different enzymes, Takara Ex Premier demonstrated results that closely mirror those obtained from PCR-free datasets, indicating its reliability in producing accurate amplifications.

how do i register a business account?
To register a business account, you can sign up directly on the website. If you need assistance during the process, it's recommended to reach out to customer support at the provided email address for help. They are available to assist with any questions or issues you may encounter while creating your account.

howdy bakhyt original question retype needed
It seems that there is a request for clarification or rephrasing of an original question, but the specific content of that question isn't provided. To assist effectively, please provide the details or context of the original query you would like to be addressed or retyped. This will help ensure a clear and accurate response.

difference def-cs def cs system single-cell single cell?
The conversation provided does not directly address the specific differences between the def-cs system and single-cell systems. However, generally speaking, a def-cs system may refer to a more integrated or comprehensive approach compared to a single-cell system, which typically focuses on individual cell analysis. The "def" in def-cs might indicate a specific feature or enhancement aimed at improving functionality or user experience, possibly for those involved in editing processes. For precise details about the differences, it would be best to consult technical specifications or product documentation related to both systems.

what can you tell me about the lenti promo and how it affects my current inventory and expiration dates?
The current inventory consists of one lot that is valid until June 24, 2025. While there are several units available within this lot, information about upcoming lots has not yet been provided. It’s a good idea to reach out to customer service for specific details regarding the lenti promo and how it may impact your inventory or future expiration dates. They will be able to provide comprehensive insights into any changes or updates related to the promotion.

is this kit suitable for both PCR clean-up and agarose gel DNA extraction?
Yes, the NucleoSpin Gel and PCR Clean-up kit is designed to be suitable for both PCR clean-up and agarose gel DNA extraction. It effectively purifies DNA from PCR reactions as well as from agarose gels, making it a versatile choice for nucleic acid purification needs.

what options are available to simplify obtaining clones using stellar competent cells and lentiviral or aav vectors?
Stellar Competent Cells are an excellent option for simplifying the cloning process, especially when working with lentiviral or AAV vectors that contain reiterated sequences. These cells have been specifically validated for the cloning and amplification of large vectors and those with repeat sequences, such as Long Terminal Repeats (LTRs) in retroviral/lentiviral constructs and Inverted Terminal Repeats (ITRs) in adenoviral vectors. Using Stellar Competent Cells included in In-Fusion Cloning kits can enhance your cloning efficiency due to their high competency levels (no less than 10^8 cfu/µg supercoiled DNA). This ensures robust transformation rates, which is particularly beneficial when dealing with complex vector systems. Additionally, employing techniques like In-Fusion Cloning streamlines the process by allowing direct assembly of fragments without the need for restriction enzymes or ligation steps, further simplifying obtaining clones. If you have any more questions about optimizing your cloning experiments or other related topics, feel free to ask!

need to add rox reference dye in syber mix for lenti-x titration kit qrt-pcr?
The mix contains TB Green, which has similar spectral properties to SYBR Green. Therefore, you can use the SYBR settings without needing to add an additional ROX reference dye in the SYBR mix for your Lenti-X titration kit qRT-PCR. It’s always good practice to double-check specific protocol details or guidelines provided with your kit for any updates or recommendations.

ever compared tested protoscript ii reverse transcriptase with neb options?
There hasn't been any specific comparison mentioned between Protoscript II Reverse Transcriptase and NEB options in the conversation. However, it is noted that some reverse transcriptases may have varying tolerances to 4sU, which can disrupt RT reactions. If you're considering a comparison, it might be worthwhile to check the compatibility of both with modified nucleotides like 4sU based on their respective product information or user experiences.

what is the recommended number of freeze thaw cycles for my stock solution?
It seems that evaporation is more of a concern for your stock solution rather than freeze-thaw cycles, as the compound is very stable. While specific recommendations can vary based on the particular substance and its stability, it's generally advisable to minimize freeze-thaw cycles when possible. If you are aliquoting into smaller volumes (like 5-use portions), this should help reduce the number of times any single aliquot is frozen and thawed, preserving its integrity over time.

what can i do to increase transduction efficiency in my mouse salivary gland organoids using plvx lentiviruses?
To increase transduction efficiency in mouse salivary gland organoids using plvx lentiviruses, consider the following strategies: 1. **Use of RetroNectin**: Since RetroNectin has been shown to enhance transduction efficiency for some cell types, you might want to experiment with T100A RetroNectin® Recombinant Human Fibronectin Fragment. This could improve the interaction between the lentivirus and your cells if they express VLA-4 and VLA-5. 2. **Optimize MOI**: Ensure that you are using an appropriate multiplicity of infection (MOI). A higher MOI can lead to increased expression levels; however, be cautious as too high a titer may cause cytotoxicity. 3. **Transduction Enhancers**: Consider incorporating transduction enhancers like Lenti-X Transduction Sponge which is designed to boost the overall efficiency of viral delivery into various cell types. 4. **Polyethylene Glycol (PEG) or Polybrene**: If you've had success with polybrene previously, it may be worth continuing its use or optimizing its concentration during transductions. 5. **Promoter Selection**: Choose strong promoters that are less likely to be silenced in your specific cell type; EF1alpha is a good option for stable expression in hematopoietic cells but evaluate others suitable for salivary gland organoids as well. 6. **Multiple Infections**: Conduct multiple rounds of infections; this can result in multiple integrations leading potentially to higher expression levels of your gene of interest (GOI). 7. **Cell Condition Optimization**: Make sure your organoids are healthy and at optimal density before performing transductions, as their condition can significantly influence uptake efficiency. 8. **Time Between Experiments**: Keep track of virus titers over time, as changes might affect outcomes on different days—using freshly prepared stocks when possible is advisable. These approaches should help maximize the effectiveness of your lentiviral transductions in mouse salivary gland organoids while also considering any unique characteristics associated with these cells.

what's the range concentration of lentivirus that can be detected?
The detection threshold for the Lenti-X GoStix Plus corresponds to a titer of approximately 5x10^5 IFU/ml. This means that lentivirus concentrations at or above this level can be detected effectively.

can you explain how to calculate the final titer of aav particles after performing real-time pcr using different dilution factors?
To calculate the final titer of AAV particles after performing real-time PCR with different dilution factors, follow these steps: 1. **Obtain Results from Standard Curve**: Use the standard curve generated during your qPCR to determine the concentration of copies per microliter (copy/µl) for your diluted sample. 2. **Account for Dilution Factors**: - If you are using a diluted sample and have included additional dilutions in your procedure, factor those into your calculations. - For example, if you took 2 µl of virus preparation and added it to 20 µl of DNase I treatment followed by another 20 µl of lysis buffer, this results in an initial dilution step. 3. **Calculate Concentration from Diluted Sample**: - Assume that you obtained a result from qPCR indicating a concentration (e.g., 6.3e4 copy/µl). - This value corresponds to the volume used in the assay (5 µl). The same concentration applies to your diluted sample. 4. **Adjust for Further Dilutions**: - If there was at least a 50-fold dilution made with EASY Dilution but only performed a specific amount resulting in a total effective dilution factor (for instance, if it turned out to be around 10), multiply accordingly. - In our example where no additional lysis buffer was added beyond what is mentioned, multiply by the appropriate factor; here it would be multiplying by either 10 or more based on actual conditions applied. 5. **Final Calculation**: - Using our earlier calculation: if you determined that there were approximately \(6.3 \times 10^4\) copies/µl in dilute form and then adjusted considering any further dilutions or concentrations backtracked through previous steps leads us to find that undiluted stock remains consistent at \(6.3 \times 10^5\) copy/µl before applying any further adjustments for volumes taken into account during sampling procedures. By systematically applying these factors—initial quantification via standard curves, accounting for all dilutions made throughout processing—you can accurately determine final titers needed for experimental applications such as retro-orbital injections or other methodologies requiring precise viral genome data input.

what could be causing the non visible pellet after centrifugation?
The non-visible pellet after centrifugation could be due to several factors. One possibility is that the concentration of the virus in the sample is low, making it difficult to visualize a distinct pellet. Additionally, if you're using low protein media, the absence of proteins like albumin can result in an invisible pellet since these proteins aid in making pellets more visible during centrifugation. It's also important to ensure that proper techniques are used during aspiration or pouring off supernatant; any disturbance could dislodge a small or less visible pellet. Marking the tube before spinning and being mindful while aspirating can help minimize this issue and ensure you capture as much of the virus as possible even if it's not visually apparent.

how to calculate final concentration of virus from ct values using lenti-x qrt-pcr titration kit?
To calculate the final concentration of virus from Ct values using the Lenti-X qRT-PCR Titration Kit, you will need to follow these steps: 1. **Standard Curve Preparation**: First, generate a standard curve using known concentrations of viral RNA or DNA. This involves running serial dilutions of your virus on the same plate as your samples and recording their corresponding Ct values. 2. **Plotting Standard Curve**: Use a graphing tool or software to plot the log of viral concentrations against their respective Ct values. Fit a linear regression line through these points to create your standard curve. 3. **Determine Sample Concentration**: Once you have established the standard curve, input your sample's Ct value into the equation derived from your linear regression (typically in the form y = mx + b). Solve for x (the log concentration). 4. **Convert Log Concentration**: If necessary, convert this log concentration back to its original form (exponential) to find out how many copies per unit volume are present in your sample. 5. **Calculate Final Concentration**: Adjust based on any dilution factors used during preparation if applicable, ensuring that you account for all steps leading up to this calculation. 6. **Reporting Results**: Finally, report your results in appropriate units such as copies/mL or another relevant measure depending on what is required for interpretation in context. Make sure all calculations consider any specific details provided with the kit regarding sensitivity and efficiency adjustments that may be needed based on experimental conditions.

can you help me with the calculations needed for determining the final titer and virus count from my real-time PCR data?
To determine the final titer and virus count from your real-time PCR data, follow these steps: 1. **Standard Curve Data**: Start with the copy number you obtained from your standard curve. In this case, it's 6.3e4 copies/µL for the diluted sample. 2. **Dilution Factors**: - The diluted sample was prepared by taking 2 µL of the virus preparation into a total volume of 60 µL (20 µL DNase I treatment + 20 µL Lysis buffer). This results in a dilution factor of 30 (since you are working with a total volume that is larger than what was initially used). - If no additional lysis buffer was added to equalize volumes as mentioned, you'd multiply by an additional factor based on how much lysis buffer would have been included. 3. **Calculation Steps**: - For the undiluted concentration: \[ \text{Concentration} = \text{diluted concentration} \times \text{dilution factor} \] Assuming no extra lysis buffer added, use a multiplication factor of 10 if only considering that step. - Therefore: Undiluted concentration = \(6.3e4\) copies/µL * \(30\) (considering there’s still some residual effect since not all buffers were accounted for). 4. **Final Titer Calculation**: Once you get your undiluted concentration in copies/µL, remember to adjust for any further dilutions made during assay setup. By following these steps carefully and applying each dilution appropriately, you'll arrive at your final titer and overall virus count accurately reflecting your PCR results.

where is my email with login details?
If you haven't received your email with the login details, it may be helpful to check your spam or junk folder. If it's not there, reaching out to the designated support team for assistance would be a good next step. They can help you retrieve your password and provide any additional information regarding discounts that might apply to your organization.

What is the recommended annealing temperature for this product?
To determine the recommended annealing temperature for the product, you first need to identify the melting temperature (Tm) of the primers being used. The manual indicates that the annealing temperature should be set at 5 degrees Celsius lower than this melting temperature. Therefore, once you have obtained or calculated the Tm of your specific primers from either their sequences or experimental data, subtracting 5 degrees will give you the appropriate annealing temperature. For precise values related to specific products mentioned in the user manual, consulting that document directly would provide more tailored information.

what's different?
The differences between EASY Dilution #9160 and EASY Dilution II #9451 primarily lie in their composition. While both products have changed, there is no significant difference in performance. EASY Dilution II #9451 is designed to accommodate a broader range of reagents. If you're currently using EASY Dilution #9160 and encounter compatibility issues with certain reagents, it may be beneficial to try EASY Dilution II #9451. However, for systems already utilizing EASY Dilution #9160, it's recommended not to switch unless you first verify the reaction within your measurement system when transitioning to EASY Dilution II #9451.

could you provide the protocol for using the AAV real-time PCR titration kit without starting from cell culture?
To use the AAV real-time PCR titration kit without starting from cell culture, you'll need to follow a protocol that begins with purified AAV viral particles. Ensure you have already purified the viral particles before proceeding. 1. Start by extracting the AAV vector genome using VI-2 of your specific protocol. 2. To release the viral DNA, make sure to include any necessary steps for lysis or extraction as outlined in your protocol. 3. Once you've released the viral DNA, you can proceed with setting up your real-time PCR according to the instructions provided with your titration kit. If you require further assistance or specific details about each step, consider reaching out to customer support for additional guidance tailored to your needs.

what's the best buffer composition to maintain protein stability during the load step?
To maintain protein stability during the load step, a good buffer composition typically includes a phosphate or Tris buffer at physiological pH (around 7.2 to 7.4). It's also beneficial to consider adding 0.15-0.3 M sodium chloride to help stabilize proteins by reducing non-specific interactions and promoting solubility. Additionally, incorporating stabilizing agents such as glycerol or sucrose can provide further protection against denaturation and aggregation during this phase. Always verify specific conditions based on your protein's characteristics for optimal results.

what is the transfer plasmid that fits my cloning project needs?
The Lenti-X™ Bicistronic Expression System (Puro), specifically the pLVX-IRES-Puro vector, is an excellent choice for cloning projects that require co-expression of two genes. This system allows for the simultaneous expression of a gene of interest and a selectable marker using an internal ribosome entry site (IRES). The provided links contain both the vector sequence and detailed information about its features and applications, which can help you determine if it fits your specific needs for gene expression or functional studies in your project. If you have more specific requirements or questions about your cloning project, feel free to ask!

possible bottle may contaminated not?
While contamination issues for this product are rarely reported, it's always a good idea to check for any signs of contamination. Warming the bottle in a 37°C water bath may help reduce thickness and clarify the contents. Additionally, examining it under a microscope could provide further insights. Keeping an eye on the outcome will be helpful in determining if there are any concerns with the bottle.

how can I minimize secondary structures when designing primers for multiple fragment cloning?
To minimize secondary structures when designing primers for multiple fragment cloning, consider the following strategies: 1. **Avoid Repeats**: Design your primers to avoid long stretches of identical nucleotides or repetitive sequences, which can promote hairpin formation. 2. **Check Melting Temperature (Tm)**: Ensure that the Tm values of your primers are within a similar range to facilitate effective annealing during PCR while avoiding conditions that might allow for secondary structures to stabilize. 3. **Use Software Tools**: Utilize primer design software that predicts potential secondary structures and provides feedback on the stability of the designed primers. 4. **Optimize Primer Length**: Aim for 20 bases at the 5' end for overlaps with adjacent sequences and keep 18-24 bases at the 3' end targeted towards annealing with your DNA template, ensuring you maintain balance in length without promoting structure formation. 5. **Temperature Considerations**: Remember that conducting PCR reactions at higher temperatures can help melt away any existing secondary structures, but it’s essential to ensure that these temperatures do not hinder proper amplification. By carefully considering these factors during primer design, you can effectively reduce potential issues related to secondary structures in your multi-fragment cloning experiments.

possible initially band develop later?
It is possible for the bands to develop later, although it is not typical. Usually, if there are delays in band appearance, it may indicate low virus production or a weaker signal that might not be visible to the naked eye but could still register on the app. If you notice any initial bands appearing after some time, it's worth monitoring their intensity over time as well. Please continue with your testing using the controls provided and let us know how that goes.

set qpcr probe based reaction page?
To set up a qPCR probe-based reaction, you'll want to refer to the user manual for the RR390A-Premix Ex Taq™ (Probe qPCR) kit. This document will provide detailed instructions on how to prepare and run your assay. You can find the user manual at this link: https://www.takarabio.com/products/real-time-pcr/great-value-master-mixes/probe-based-mixes/premix-ex-taq-(probe-qpcr)?catalog=RR390A. Make sure you have already generated your cDNA using the RR047A kit, as it is specifically designed for creating cDNA suitable for qPCR applications. If you have any further questions or need additional assistance with your setup, feel free to ask!

how can i place an order without an account?
To place an order without an account, you will need to contact customer service directly. They can assist you in setting up an account and processing your order. You can reach out to them via phone or email for assistance with the registration process and placing your order.

how can i successfully create a research account with takara?
To successfully create a research account with Takara, you should visit their registration page and follow the prompts to sign up. If you encounter any issues, such as your email already being in use, it may be necessary to reach out for assistance. You can contact their support team at the designated email address for help with setting up your account. Once your research account is established, you will then be able to proceed with creating a business purchasing account if needed.

can you provide the details on how to generate a stable cell line with puro resistance?
To generate a stable cell line with puromycin (Puro) resistance, you can utilize a monocistronic vector that expresses the Puro resistance gene directly. This approach simplifies the process as it avoids complications associated with IRES constructs and potential issues related to open reading frames. Here are the steps to follow: 1. **Choose a Monocistronic Vector**: Select or design a vector that contains the Puro resistance gene under a suitable promoter for your target cells. 2. **Transfect Cells**: Introduce this vector into your target cells using an appropriate transfection method (such as lipofection, electroporation, or viral transduction). 3. **Selection of Resistant Cells**: After transfection, expose the cells to puromycin at selected concentrations over several days to kill off non-resistant cells. 4. **Isolation of Surviving Clones**: The surviving colonies will contain your stably integrated Puro-resistant cells. You can expand these colonies for further analysis or experimentation. If you're considering alternative methods like lentiviral vectors, those could also be used but might involve additional complexity in terms of packaging and delivery systems. Letting some sequences serve as stuffing material in case you're dealing with IRES constructs may help mitigate potential issues if that's still on your mind; however, focusing solely on integrating Puro through direct expression is often more straightforward for achieving stability in resistant cell lines.

what type of membrane used different mwco spin columns?
Different molecular weight cut-off (MWCO) spin columns typically use membranes made from materials like polyethersulfone (PES), regenerated cellulose, or nylon. The choice of membrane material can influence the separation efficiency and compatibility with various samples. Each type of membrane has specific characteristics regarding protein binding, flow rates, and chemical resistance, which are important to consider based on your purification needs. If you have a particular application in mind, it may be helpful to refer to product specifications for more detailed information related to MWCO options.

Can I still use the lysis and neutralization buffers if they're no longer sold in liter bottles?
Yes, you can still use the lysis and neutralization buffers even if they're no longer sold in liter bottles. As long as you have access to the appropriate volumes of these buffers from other available sizes or sets, they will work effectively for your applications. It's important to ensure that you're using the correct protocol and concentrations as specified in your experiments. If needed, consider ordering smaller quantities or buffer sets that include these components.

what's status first run kits order package replacements
The status of the first run kits order package replacements would typically depend on your specific order details and any communications you've had with your supplier. It's best to reach out directly to the customer service or sales representative associated with your account for updates regarding replacements or issues related to your kit order. They will have the most accurate information regarding availability, timelines, and any potential solutions. If you need assistance in contacting them, let me know!

recommend sequence for stuffing that won't affect cellular function?
For stuffing sequences that won't affect cellular function, it's generally a good idea to use well-characterized and non-disruptive sequences. Common options include: 1. **Linker Sequences**: Short peptide linkers (e.g., Gly-Ser repeats) can be inserted as they are typically neutral and do not interfere with protein folding or function. 2. **Fluorescent Protein Genes**: Using a small fluorescent protein like GFP or mCherry can serve both as a visual marker and a functional filler without significantly impacting the downstream ORF expression. 3. **Common Reporter Genes**: Small reporter genes such as luciferase can also work effectively if you aim for dual-functionality in your construct. 4. **Synthetic Untranslated Regions (UTRs)**: Incorporating synthetic UTRs that do not code for any proteins but help maintain structural integrity could be beneficial. It's important to ensure that whatever sequence is chosen does not introduce unwanted regulatory elements or secondary structures that could interfere with translation efficiency of the IRES-driven ORF. Testing different constructs will ultimately provide insights into what works best for your specific application without compromising cell functionality.

how do I design primers for In-Fusion cloning to replace a long stretch of DNA with a smaller affinity tag while considering linearization and divergent primers?
To design primers for In-Fusion cloning aimed at replacing a long stretch of DNA with a smaller affinity tag, while considering linearization and the use of divergent primers, follow these steps: 1. **Identify the Target Region**: Determine the specific sequence in your plasmid that you want to replace with your affinity tag. 2. **Design Divergent Primers**: Create two primers that flank the region to be replaced. These primers should be designed to anneal to sequences immediately adjacent (but outside) of the target area. - The 5' end of each primer should include additional nucleotides corresponding to your desired insert (the affinity tag) along with any necessary restriction sites if applicable. 3. **Add Homology Regions**: For In-Fusion cloning, ensure that both primers have complementary ends matching the sequence of your insert. Typically, this means including about 15-20 bp homology regions at their 3' ends that are homologous to the adjacent vector sequences where you want them integrated. 4. **Linearize Your Vector**: If you're using PCR or restriction enzyme digestion for linearization: - Ensure that this process does not cut within your intended insertion site but creates an open circular structure suitable for inserting your new fragment. - If using PCR for linearization, design internal oligonucleotides as needed based on template orientation. 5. **Check Specificity and Melting Temperature (Tm)**: Validate primer specificity against your vector's entire sequence using tools like BLAST and ensure they have similar Tm values (ideally within 2-3°C). 6. **Order and Prepare Your Insert**: Once you've confirmed optimal designs through simulations or software analysis, synthesize or clone your affinity tag into a separate vector if required before proceeding with In-Fusion assembly. 7. **Confirm Cloning Strategy Compatibility**: After sequencing verification post-cloning, confirm successful integration by analyzing via PCR or restriction digest methods targeting around insertion points. By following these guidelines carefully while designing divergent primers tailored specifically towards facilitating efficient In-Fusion cloning will enhance success rates in achieving precise genetic modifications such as integrating an affinity tag seamlessly into existing plasmids without disrupting crucial downstream elements essential for expression studies or assays later on.

what is seek curiosity lmp?
Seek Curiosity LMP refers to a specific workflow or method used in laboratory settings, particularly in relation to the analysis of data and results. In this context, "LMP" typically stands for "lamp," which indicates that it may involve protocols associated with LED-based light sources, as mentioned in your conversation. This approach encourages exploration and inquiry within scientific processes, aiming to enhance understanding and innovation by leveraging existing technologies like those found in systems such as the QuantStudio 7 or ABI7500 lines.

know volume needed for cleancap to compare costs effectively?
To determine the volume needed for CleanCap AG to compare costs effectively, you can use the concentration information provided. Since the CleanCap AG is supplied at a concentration of 100 mM and each tube contains 1 µmol (N-7113-1), you can calculate the required volume based on your desired final concentration in your experiments. For example, if you want to prepare a solution with a certain molarity using this stock solution, you'll need to know how much of that stock solution corresponds to your target amount. For instance: - If you're preparing a total of 10 µmol for an experiment, you'd take 100 µL from the N-7113-1 stock since: \[ Volume = \frac{Desired\ Amount}{Concentration} = \frac{10\ µmol}{100\ mM} = 0.1\ mL\ or\ 100µL \] You can adjust this calculation based on how much of each reagent you plan to use in your assays or experiments. This way, you'll be able to compare costs more accurately by knowing exactly how much CleanCap AG you will require for different applications. If there's anything specific you'd like further clarity on regarding volumes or calculations, feel free to ask!

where is my order for the titer virus?
It seems that the titer information is being checked internally, and you will be updated via email once it is available. If you have any other questions or need further assistance, feel free to ask. Thank you for your patience!

what kit should i buy for amplifying lncrna from my cDNA samples?
For amplifying lncRNA from your cDNA samples, you should consider using the RACE kit, specifically designed for 5' and 3' RACE (Rapid Amplification of cDNA Ends). This kit includes universal primers that will help in selecting the ends of your RNA. It's important to ensure that your cDNA recognizes these universal primers for effective PCR amplification. The user manual provides detailed information on how the specific primers target mRNA and include sequences that are compatible with the universal primer mix. You can start directly from RNA as indicated in the protocol.

what works that's?
The conversation indicates that the Discount Quote Q106603 is important for receiving a discount when placing an order. If there are any issues with using the quote, it's advised to contact customer support for assistance. If you have further questions or need additional help, feel free to ask.

what could be the reasons for gibson assembly not working despite using overlapping primers?
There could be several reasons why Gibson assembly might not work even when using overlapping primers. One common issue is the difference in how overlap regions are calculated for various cloning methods, such as In-Fusion and Gibson assembly. If the overlap sequence isn't correctly designed according to the specific requirements of Gibson assembly, it can lead to incorrect annealing or failure to join fragments. Another potential reason could be that the lengths of the overlaps may not be optimal; typically, a minimum of 20-40 base pairs is recommended for effective joining. Additionally, if there are secondary structures within the DNA fragments or primer-dimer formations, these can hinder proper assembly. The quality and concentration of your DNA fragments also play a crucial role—degraded or low-concentration templates can affect success rates. Furthermore, ensuring that all components of your reaction mix are fresh and prepared correctly is essential for efficient assembly. If you encounter ongoing issues despite checking these factors, reviewing detailed protocols and consulting user manuals specific to your reagents may provide further insights into troubleshooting steps.

where is my quote for the single plate?
The quote for the single plate is currently being prepared and should be sent to you soon. If there are any specific details or questions you have about the project or other needs, feel free to let me know!

what should i do about the room temperature virus package?
It’s important to assess the situation carefully. If the package containing the virus was kept at room temperature for at least a day, there may be concerns about its viability. It would be advisable to follow your organization's protocols regarding handling potentially compromised materials. Additionally, consider consulting with a supervisor or relevant authority to determine the best course of action and whether further testing or disposal is required. If you need assistance in tracking down delivery details or confirming any specifics, gathering all necessary order information will help facilitate that process.

what is the recommended unit of coli dna ligase enzyme per reaction and what is the protocol for using it in a specific reaction volume?
The recommended unit of the coli DNA ligase enzyme is typically based on the specific requirements of your experiment, but a common guideline suggests using 1-2 units per reaction, depending on the amount of DNA and desired efficiency. For example, if you are working with λ DNA-Hind III fragments in a 20 μl reaction volume, you could use approximately 1-2 units of the enzyme. Here’s a general protocol for using it: 1. Prepare your reaction mixture by combining: - The appropriate amount of DNA (e.g., λ DNA-Hind III fragments). - Buffer suitable for ligation (often included in commercial preparations). - Enzyme: Add 1-2 µl (equivalent to 60-120 U) from the stock solution. - Adjust final volume to 20 μl with nuclease-free water. 2. Mix gently by pipetting up and down or flicking the tube. 3. Incubate at 16°C for about 30 minutes to allow ligation to occur. 4. After incubation, proceed with downstream applications such as transformation or analysis as needed. Always refer to specific product recommendations and adjust conditions based on your experimental needs.

which smart-seq kit is best for my scrnaseq experiment?
The choice of the best SMART-Seq kit for your single-cell RNA sequencing (scRNA-seq) experiment depends on your specific needs. If you are working with 1 to 1,000 whole cells, the SMART-Seq® mRNA Single Cell kit is a great option as it is specifically designed for this purpose. Additionally, if you're looking for low input requirements, the renamed SMART-Seq® v4 Ultra® Low Input RNA Kit might also be suitable. For library construction and dual indexing, consider using the SMART-Seq® mRNA Single Cell LP kit along with the Unique Dual Index Kit that you'll need to purchase separately. To make an informed decision based on sensitivity and performance, refer to the technical notes which compare various kits available. If you'd like more detailed assistance tailored to your experimental design or any other questions arise later, reaching out through official support channels can provide additional guidance.

what's typical yield fusion needs ug cloned product?
The typical yield for In-Fusion cloning can vary depending on several factors, including the complexity of the insert and vector, as well as the efficiency of your transformation method. Generally, users may expect yields in the range of 1 to 10 micrograms (µg) of cloned product after performing In-Fusion cloning. It's important to optimize conditions based on your specific experiment for best results. If you have further questions about optimizing yield or any other related topics, feel free to ask!

what's typical yield for fusion and can we amplify directly without transformation?
The typical yield for In-Fusion cloning can vary depending on the specific conditions and the DNA involved, but it generally provides high efficiency in generating correctly ligated plasmids. However, direct amplification of In-Fusion products without transformation into bacteria is not feasible because the resulting DNA is not a "closed" plasmid; it's linearized and needs to be circularized through transformation first. This means that you cannot amplify directly from an In-Fusion product before transforming it into competent cells. If you're looking for alternative methods, consider contacting your project manager for more insights on upcoming products that may suit your needs better.

coa corresponds to which lot number received?
The Certificate of Analysis (CoA) typically corresponds to a specific lot number associated with the product. To determine which lot number is linked to the CoA you referenced, it would be best to consult your quality assurance team or check the documentation that came with the product. They should be able to provide you with the exact lot number related to that CoA.

how can i reach hr austin takar to inquire about my application?
To inquire about your application, it's best to wait for communication from the HR department, as they handle the selection process and will provide updates on your status. If you have specific inquiries related to a job application, consider checking if there are any contact details provided in the job listing or on the company's careers page. Unfortunately, direct access to HR contacts isn't available at this moment.

guys sell unstained version mcherry flow beads
It seems that there are no unstained versions of the mCherry flow beads available for sale. However, you may be able to purchase the beads used for coating with mCherry or AcGFP1. If you're looking for specific details about these beads, I can check with our R&D team and see if we can provide more information. Let me know if there's anything else you need!

what are the best recommended takara indexes to use for picoplex gold kit step amplification?
For the Picoplex Gold kit step amplification, it is recommended to use Takara's full-length barcoded adapters. Specifically, ensure that you select compatible indexes like i7 and i5 that match your sequencing needs. The provided sequences for Index 1 (i7) and Index 2 (i5) can serve as a reference when choosing suitable Takara adapters. Additionally, keep in mind that due to variations in primer concentrations, optimizing PCR cycling conditions may be necessary for optimal results.

how can I place an order without having my own account?
You can place an order without having your own account by calling customer support directly at the provided phone number. They are available Monday through Friday from 6:00 am to 5:00 pm Pacific time. Alternatively, you can also reach out via email for assistance with placing your order. If you have any further questions or need additional help, feel free to ask!

how to transform plasmid containing ccdb gene into normal e-coli strain using stellar competent cells with a compatible plasmid?
To transform a plasmid containing the ccdb gene into a normal E. coli strain using Stellar competent cells, follow these steps: 1. **Prepare Competent Cells**: Ensure that you have Stellar competent cells ready for transformation. These should be thawed on ice and kept cold throughout the process. 2. **Mix Plasmid with Competent Cells**: In a sterile tube, combine an appropriate amount (usually 50-100 ng) of your plasmid DNA with the prepared Stellar competent cells (typically around 50 µl). Gently flick the tube to mix without creating bubbles. 3. **Incubate on Ice**: Let this mixture sit on ice for about 30 minutes to allow for optimal uptake of DNA by the competent cells. 4. **Heat Shock Treatment**: Perform a heat shock by placing the tube in a water bath at 42°C for approximately 45-60 seconds. This step is crucial as it helps facilitate the entry of plasmid DNA into the bacteria. 5. **Recovery Phase**: Immediately return the tubes to ice for another 2 minutes, then add SOC or LB medium (without antibiotics) to help recover and express any antibiotic resistance genes present in your plasmid—this typically involves adding around 950 µl of medium. 6. **Incubate at Optimal Temperature**: Allow this mixture to recover by incubating it at 37°C for about one hour with shaking (if possible). 7. **Plating Transformants**: After recovery, plate an appropriate volume onto agar plates containing selective media that corresponds with your plasmid's selection marker(s). For example, if your plasmid has an ampicillin resistance gene, use LB agar supplemented with ampicillin. 8. **Incubation of Plates**: Incubate these plates overnight at suitable growth conditions (usually at 37°C). 9. **Screening Colonies**: Following incubation, look for colonies that have grown; these will contain your transformed E.coli expressing both viable characteristics from Stellar and possibly carrying other genes like ccdb depending on selection methods used during transformation. By following these steps carefully while ensuring all materials are sterile and reactions are performed optimally, you can successfully transform E.coli strains using Stellar competent cells despite initial concerns regarding resistance traits associated with toxic genes like ccdb.

where is my order for the gummis?
It seems that there may have been some difficulty locating your order. To assist you better, please provide your order number or purchase order (PO) number. If you don't have this information, it would be best to directly contact the orders department for further assistance. They can be reached at 800-662-2566, option 1, from Monday through Friday between 6:00 am and 5:00 pm Pacific time. Alternatively, you can also reach out via email if needed.

what additional stability data can you provide for single shot lots to address the efficacy concerns?
To address the efficacy concerns regarding the stability of single shot lots, it's important to note that under proper storage conditions—specifically being desiccated and protected from moisture—the single shot should remain stable over an extended period. The shelf life is indicated as one year in the Certificate of Analysis, which aligns with our R&D findings. They found it challenging to reproduce failure scenarios when using the single shot, suggesting robust performance under typical conditions. Additionally, minimizing the time between preparation of the polymer/plasmid mix and its application to cells can help enhance effectiveness. While temperature changes are not a significant concern for this reagent, exposure to moisture should be avoided to maintain stability. If more detailed internal data is required, I will coordinate with team members who can provide further insights or conduct additional testing if necessary.

wondering what low input kits with umis are similar to thruplex dna-seq kit?
The ThruPLEX® DNA-Seq Kit is not compatible with UMI adapters, as the UMI sequences are incorporated during the Stem-Loop adapter ligation step before using the Indexing adapters. For low input kits that do support UMIs, you might consider exploring Tag Seq kits; however, please note that they typically require a higher input amount than ThruPLEX. Further information may be needed to identify specific alternatives that fit your requirements.

know new product size difference that's bit smaller version similar product?
The smaller version of the adenovirus purification kit, which you referred to as 631532 (2 preps), is being discontinued. The available option is a larger size kit, 631533, which still serves a similar purpose but offers more preparations. For further details on product sizes and specifications, please check the manufacturer's website directly.

why is my research account not letting me purchase items online?
It sounds like you may need to upgrade your research account to a purchasing account. Typically, after signing up for a research account, there is an additional form that needs to be completed online for the purchasing account. It would be best to reach out directly to customer support for assistance with this process, as they can provide specific guidance tailored to your situation.

anyone ever tried using magnetic beads for viral transduction in early embryos?
The conversation does not explicitly mention any attempts to use magnetic beads for viral transduction in early embryos. However, it discusses the application of magnetic beads and their effectiveness with various viruses, including lentiviruses and MMLV. While there is interest in optimizing their use, particularly regarding incubation times and bead quantities, no specific references or experiences related to early embryos are noted. It may be beneficial to explore literature on similar applications or reach out to researchers who specialize in this area for more targeted insights.

can you provide transduction efficiency data for the lentivirus products?
Transduction efficiency data for lentivirus products can vary based on several factors, including the specific cell type being targeted and the conditions used during transduction. While there may not be direct data available in your current resources, it's a good idea to check published literature related to your lentivirus system or consult with colleagues who might have conducted similar experiments. If you're looking specifically for efficiencies related to colon cancer cells or EMT-6 mammary tumor cells, examining studies that focus on those models could provide useful insights. Additionally, if RetroNectin is being utilized primarily for hematopoietic cells, it would be beneficial to verify whether its application is relevant in your context of generating CAR-T cells against mouse colon or mammary tumors. Gathering this information will help clarify the potential effectiveness of your gene transfer efforts using the lentiviral approach.

what could be causing the undetermined pcr results even though it worked previously?
Undetermined PCR results, especially when the assay worked previously, could be caused by several factors. It might indicate that your sample is below the detection limit of the assay. Other possible reasons include issues with reagent quality or concentration, such as improper handling or expiration of components like ROX dye or the premix itself. Additionally, variations in template quality or quantity can affect amplification. It's also important to ensure that all thermocycler settings are correct and compatible with your specific protocol. If you have recently changed any reagents or protocols, that may also contribute to this issue.

are stellar competent cells from clontech labs recommended for in-fusion assembly work?
Stellar competent cells from Clontech Labs are indeed recommended for In-Fusion assembly work. They have been optimized for high transformation efficiency, which is essential for the success of cloning and assembly techniques such as In-Fusion. It's always a good practice to refer to the specific product documentation and any available certificates of analysis to ensure compatibility with your experimental needs.

how to efficiently transfect mammalian cells with two sgRNA plasmids for gene knockout?
To efficiently transfect mammalian cells with two sgRNA plasmids for gene knockout, you can follow these steps: 1. **Cotransfection**: Prepare and cotransfect the two sgRNA plasmids into your target mammalian cells using an appropriate transfection reagent that is compatible with your cell type. This allows both sgRNAs to be expressed simultaneously in the same cells. 2. **Screening for Knockouts**: After transfection, it’s essential to screen the cells to confirm successful knockouts as multiple sgRNAs may lead to off-target effects or varied efficiencies. 3. **Cloning Two sgRNA Cassettes**: If you prefer a single vector approach, consider cloning two distinct sgRNA expression cassettes into a single plasmid if your vectors allow for this modification. This method could simplify the process and improve co-expression of both guide RNAs. 4. **Use Appropriate Vectors**: Make sure you are using suitable vectors that support dual expression of sgRNAs if going down this route; some commercial options might offer such capabilities. 5. **Consult Technical Resources**: Review available technical notes and resources from reputable sources such as Takara Bio or Addgene that provide guidance on cloning strategies and protocols specific to CRISPR/Cas9 applications in mammalian systems. By employing these approaches, you'll increase the likelihood of achieving effective gene knockout through simultaneous targeting with multiple sgRNAs.

is it possible to skip adding indexes during amplification step and still use the amplified product for downstream sequencing?
Skipping the addition of indexing adapters during the amplification step is not advisable if you plan to use the amplified product for downstream sequencing. Indexing adapters are essential for identifying and distinguishing between different samples in a sequencing run, especially when multiplexing multiple libraries. Without these indexes, it would be challenging to accurately analyze or sequence your samples as they may not be uniquely identifiable during the sequencing process. Therefore, it's recommended to include indexing adapters during amplification to ensure successful and efficient downstream applications.

what is the recommended primer length for successful mutagenesis in fusion cloning, especially considering I want to ensure complete mutagenesis and avoid missing nucleotides at the ends?
For successful mutagenesis in fusion cloning, it is recommended to use primers with an overlap length of no shorter than 13 bases and no longer than 21 bases. This range helps ensure optimum efficiency while incorporating mutations. Additionally, you have the flexibility to design sequences up to approximately 65 bases long for including mutations within your primers. Using a primer design tool, such as the one provided by Takara, can assist in creating effective primers tailored for your specific needs.

how do i reset my temporary password to place my order?
To reset your temporary password, you can typically follow the prompts on the website's login page. Look for a "Forgot Password?" link, which will guide you through the process of resetting it. If you need further assistance, feel free to call customer service at 800.662.2566 and select option 1 for help with your order and account access.

what information do I need regarding dnase treatment during human rna extractions?
During human RNA extractions, it's important to consider whether DNase treatment is performed or not. If total RNA is not treated with DNase, it may contain genomic DNA (gDNA) contamination, which can affect downstream applications such as qPCR or sequencing. It's essential to check for gDNA contamination in your RNA samples even if you do not treat them with DNase. Additionally, understanding the integrity of your RNA—whether low or high—is also crucial for ensuring quality results in your experiments. If further questions arise later regarding this topic or any other technical inquiries, reaching out to technical support would be beneficial.

what is the availability and pricing for the universal human reference rna specific tissue organ rna?
The availability and pricing for Universal Human Reference RNA, which includes specific tissue or organ RNA samples, can vary depending on the supplier. Generally, these products are available from various biotechnology companies that specialize in molecular biology reagents. It's advisable to check with suppliers like Thermo Fisher Scientific, Agilent Technologies, or similar organizations for the most current information on stock status and pricing. Additionally, it may be beneficial to contact their customer service directly for any specific inquiries regarding bulk orders or custom requests related to tissue-specific RNAs.

do you think there are specific setups or conditions we should use for in-fusion fusion ligation to improve success?
For successful In-Fusion reactions, it's crucial to adhere closely to the guidelines provided in the user manual. The setup should involve using the correct concentrations of DNA fragments and enzymes as specified. Since In-Fusion is distinct from traditional ligation methods, it’s important not to confuse the two; there’s no ligase involved in this process. Avoid extending incubation times excessively, as this may lead to unwanted digestion of your DNA. It's also beneficial to perform a comparison by running both a traditional ligation reaction and an In-Fusion reaction on a gel; you’ll observe that they yield different results due to their unique mechanisms. Ultimately, using recommended protocols without unnecessary modifications will enhance your chances of success with In-Fusion cloning.

add cdna amp cycles carry add cycles final library amp step?
To add cDNA amplification cycles in the final library amplification step, you typically follow this process: 1. **Determine Initial Conditions**: Start with your cDNA libraries that have been prepared for sequencing. 2. **Decide on Cycle Number**: Based on the concentration and quality of your cDNA, decide how many additional PCR cycles are needed. This can depend on factors such as input amount and desired yield. 3. **Set Up PCR Reaction**: Prepare a PCR reaction mix including primers specific to your library, dNTPs, buffer solution, and DNA polymerase. 4. **Add Cycles to Protocol**: Incorporate the chosen number of additional amplification cycles into your protocol. It's common to start with an initial denaturation step followed by 20-30 cycles of denaturation, annealing (at primer-specific temperatures), and extension. 5. **Final Extension Step**: After completing the necessary cycles for amplification, include a final extension step (usually around 5-10 minutes at 72°C) to ensure all fragments are fully extended. Ensure that you're monitoring yields through quantification methods like qPCR or bioanalyzer checks after each step if possible, so that you can optimize conditions based on results obtained from earlier experiments or runs.

how can i talk to someone for help with my account?
For assistance with your account, you can reach out to the designated support team via email. They are typically prompt in responding to inquiries related to account or login issues. Unfortunately, direct phone support is not available, and the tech support team does not have access to customer accounts. Following this procedure will ensure that your issue is addressed efficiently.

what is the concentration of specific fragment sizes in the ladder?
The datasheet does not provide specific information regarding the concentration of each fragment size in the ladder. It only mentions that there are 10 fragments between 20 and 200 bp in multiples of 20 bp, along with additional fragments at 300, 400, and 500 bp. Since this information is not available in the datasheet, it would be best to reach out to the manufacturer for clarification on whether they can provide details about the concentrations of specific fragment sizes.

can you provide the exact order and details for obtaining new indexed forward primers from illumina?
To obtain new indexed forward primers compatible with the SMARTer smRNA-Seq Kit for Illumina, you need to order indexing primers directly from Takara Bio USA. These proprietary indexing primers have specific overlaps that are essential for proper annealing during first-strand cDNA synthesis. The kit uses TruSeq CD (formerly known as TruSeq HT) indexes, specifically F1–F8 corresponding to D501–D508 and R1–R12 corresponding to D701–D712. You can find these details in the user manual provided by Takara Bio. If you're looking for alternative options, consider the Unique Dual Index Kit available from Takara Bio. For small RNA workflows, it's recommended to use 4 µL of each UDI primer. It's important to note that using indexing primers from Illumina or other vendors may not be compatible with the SMARTer smRNA-Seq Kit workflow due to differences in adapter sequences; therefore, make sure any alternatives have exact overlaps as required. Considering potential concerns about your current kit's age and storage conditions, it might also be wise to purchase a completely new kit if key components are missing or uncertain.

can you help me with psin plv for lentivirus production?
Yes, we can assist you with the pLV plasmid for lentivirus production. While we haven't tested all vectors, our packaging system is designed to efficiently package any HIV-1-based vector, which should yield high-titer lentivirus. We have successfully tested the ThermoFisher vector that lacks the WPRE and cPPT/CTS sequence elements and utilizes self-inactivating (SIN) LTRs, achieving over 3 x 10^7 IFU/ml. You can find more information about the vectors we've tested on our website at Takara Bio's learning center. If you need further assistance or specific details regarding your project, feel free to ask!

am wondering know antibody detect specific ecotropic envelope?
It seems there may be a gap in the availability of antibodies specifically targeting the ecotropic envelope. While you mentioned that you expected multiple antibodies validated against mCAt-1, it appears they might not be readily available on the market. It could be useful to consult with your R&D team for any insights or recommendations regarding this target, as they may have more information or alternative solutions. Additionally, exploring academic literature or reaching out to suppliers who specialize in custom antibody development could also yield some options.

what's recommended sequencing length for single read run using custom primer in the guide-it guide crispr genome-wide sgrna library protocol?
The recommended sequencing length for a single read run using the custom primer in the Guide-it™ CRISPR Genome-Wide sgRNA Library protocol is 60 bases.

how apply promo code using bearbuy system
To apply a promo code using the BearBuy system, look for a designated box during the checkout process where you can enter your discount or promo code. If you don't see it, it may be labeled as "description." If you're still having trouble finding the option to enter your code, consider reaching out to customer service for assistance. They can provide guidance on how to apply your promo effectively.

what should i do if my nanodrop readings are not working after using the cleanup kit?
If your Nanodrop readings are not working after using the cleanup kit, consider a few troubleshooting steps. First, ensure that you used the correct buffers for binding, washing, and elution during the DNA extraction process. If you suspect that the buffers may not have worked properly, try to salvage your eluted material by performing an ethanol precipitation to concentrate the DNA. This can help increase your yield and improve your readings. Additionally, double-check that you're measuring in the appropriate range for Nanodrop and that there’s no contamination on the measurement surface. If you do have access to additional buffer solutions compatible with your columns or kits, using those might also enhance results. If issues persist despite these steps and if more support is needed regarding specific products or protocols, reaching out to technical support from relevant suppliers could provide further guidance tailored to your situation.

able purchase inhibition buffer separately dna stool kit?
Lysis Buffer ST2, which is used for removing inhibitors from stool samples during DNA extraction, is not available for purchase separately. It is specifically included in the DNA stool kit to facilitate the process of clearing contaminants and ensuring high-quality DNA isolation. If you have any other questions or need assistance with a different product, feel free to ask!

would smarter universal low input kit rrna depletion zapr provide effective rRNA depletion for viral RNA in human wastewater samples?
The SMARTer Universal Low Input Kit with rRNA depletion is primarily designed for mammalian RNA and may not effectively deplete bacterial rRNA, which could impact the analysis of viral RNA in human wastewater samples. Since wastewater contains a diverse array of genetic material, including both human and microbial sources, it’s essential to consider how the kit's design aligns with your specific needs for identifying viral RNA. If you're focusing on viral RNA detection in wastewater, you may want to explore other approaches or kits specifically optimized for that purpose. While the SMARTer Stranded Total RNA-Seq Kit v3 - Pico Input Mammalian facilitates total nucleic acid extraction from various biological matrices, it's important to ensure that any method used will adequately address the complexities of your samples without significant bias towards bacterial components.

hi can you provide more details about the insect input kit for hmw dna isolation?
The insect input kit for high molecular weight (HMW) DNA isolation emphasizes the importance of effective lysis or homogenization of samples. For optimal results, it's recommended that customers use liquid nitrogen and bead beating techniques to thoroughly homogenize insect tissues. This method allows for higher sample input compared to enzymatic lysis. For processing up to 30 mg of insect samples, using NucleoSpin® Bead Tubes Type A is advised. These tubes contain 0.6–0.8-mm ceramic beads in a 2-ml tube, which facilitate effective cell disruption when used with bead beating methods. Additionally, for difficult-to-lyse samples such as insects, yeast, or bacteria, bead beating provides an advantage by allowing increased sample quantities while ensuring quality DNA extraction. The product is available at approximately USD $131.00 through Takara Bio's website. If you have any other questions regarding protocols or specific applications related to HMW DNA isolation from insect samples, feel free to ask!

what could be the reason for the lower quality of samples and how can we ensure better quality in future submissions?
Lower quality of samples can be attributed to several factors, including improper sample collection or handling, contamination during processing, or degradation over time. To ensure better quality in future submissions, consider the following recommendations: 1. **Sample Collection**: Follow standardized protocols for sample collection to minimize variability and contamination. 2. **Storage Conditions**: Maintain appropriate storage conditions (e.g., temperature and humidity) to preserve sample integrity before submission. 3. **Handling Procedures**: Ensure that all personnel involved in the handling of samples are trained on best practices to avoid introducing contaminants. 4. **Quality Control Checks**: Implement regular quality control measures throughout the sampling process to identify potential issues early on. 5. **Documentation**: Provide detailed documentation regarding how samples were collected and processed for better evaluation. By addressing these areas, you can improve the overall quality of your samples in future submissions.

what is the mechanism of ion exchange in poly-amine and how can it be incorporated into a dx platform?
The mechanism of ion exchange in poly-amine typically involves the interaction between positively charged amine groups and negatively charged ions or molecules present in a solution. This process allows for the selective binding and removal of specific anions, which can be influenced by factors such as pH, ionic strength, and the structural characteristics of the polymer. Incorporating this mechanism into a diagnostic platform could involve designing systems where poly-amine materials are used to capture target analytes through ion exchange. By optimizing conditions like buffer composition and pH, it may be possible to enhance binding efficiency and specificity for various applications, such as nucleic acid isolation or protein purification. If you need more detailed information on incorporating these concepts into a diagnostic platform, I recommend discussing with experts who specialize in material science or bioengineering to explore potential methodologies that align with your objectives.

Why is the QuantStudio machine not listing the SYBR option for Lenti-X qRT-PCR?
The absence of the SYBR option for Lenti-X qRT-PCR on the QuantStudio machine may be due to its specific configuration or intended use. Since this model is designed for digital PCR, it might not support SYBR settings unless it's dedicated to probe assays. It's advisable to check with Fisher regarding compatibility, especially in terms of whether the green channel functions effectively with SYBR or if TB Green can be used interchangeably since they share similar spectral properties. Additionally, consider that while your PCR enzyme should generally work across different machines, the premixed nature of your dye could influence performance depending on how it interacts with the system's specifications.

where can I find the protocol for using primescript rt master mix for cDNA synthesis and real-time PCR?
The protocol for using PrimeScript™ RT Master Mix (RR036A) for cDNA synthesis and real-time PCR can be found in the user manual provided at the link you shared: https://www.takarabio.com/documents/User%20Manual/RR036A/RR036A_UM.pdf. This document will contain detailed instructions and guidelines to help you with your experiments.

is there a comparison between nucleospin rna kit and qiagen rneasy maxi kit in terms of yield?
The conversation does not provide a direct comparison between the NucleoSpin® RNA Midi kit and the Qiagen RNeasy Maxi kit in terms of yield. However, it mentions that the typical yield for the NucleoSpin® RNA Midi is 180 µg from 10^7 HeLa cells and up to 620 µg from 4 x 10^7 HeLa cells. To make a thorough comparison regarding yields, specific data for the Qiagen RNeasy Maxi kit would be needed. If you're looking for detailed comparisons or performance metrics, it's advisable to consult product specifications or published studies that directly compare both kits under similar conditions.

can you send link for thruplex kit used in targeted dna-seq?
You can find information about the ThruPLEX kit for targeted DNA sequencing on the Takara Bio website under their next-generation sequencing section. This will provide you with detailed protocols and specifications related to the kit. If you're looking for specific applications or data, exploring their resources may also offer additional insights into its usage in experiments like yours.

pcr protocol shows denaturation steps and times, can you clarify the order of these steps?
The PCR protocol consists of three main steps that are repeated for a total of 30 to 35 cycles. The order of these steps is as follows: 1. **Denaturation**: This step occurs first, where the reaction mixture is heated to 98°C for approximately 10 seconds. This high temperature separates the DNA strands. 2. **Annealing**: Following denaturation, the temperature is lowered to around 62°C for about 5 seconds. During this phase, primers anneal to their complementary sequences on the single-stranded DNA templates. 3. **Extension**: After annealing, the temperature is set to 72°C for an extension time based on your product size (typically around 5-20 seconds per kilobase). This allows the DNA polymerase to synthesize new DNA strands by adding nucleotides. These three steps are then cycled through multiple times (30-35 cycles) to amplify your target sequence effectively. Adjusting each step's duration and temperatures can help optimize amplification depending on your specific conditions and template requirements.

how to use transfected cells for dna-protein crosslinking with plasmid?
To use transfected cells for DNA-protein crosslinking with plasmid, you can follow these general steps: 1. **Transfection**: Begin by transfecting your target cells with the plasmid DNA that encodes the protein of interest. Ensure that the amount of DNA used does not hinder the binding efficiency of any conjugated proteins. 2. **Expression Confirmation**: After transfection, allow sufficient time for gene expression (typically 24-48 hours). Confirm successful protein expression through techniques such as Western blotting or immunofluorescence. 3. **Crosslinking Setup**: Prepare a crosslinking solution, often using formaldehyde or another suitable crosslinker compatible with your experimental needs and cell type. Optimize concentration and incubation time to achieve effective crosslinking without excessive background noise. 4. **Treatment of Cells**: Treat the transfected cells with the crosslinker under conditions optimized for your specific experiment (e.g., temperature, duration). Be sure to quench unreacted crosslinker afterward using an appropriate quenching buffer (such as glycine). 5. **Cell Lysis**: After quenching, lyse the cells to release cellular components while maintaining protein integrity for downstream analysis. 6. **Analysis**: Proceed with purification and analysis techniques such as immunoprecipitation followed by mass spectrometry or Western blotting to investigate interactions between your protein and associated DNA. 7. **Controls**: Include necessary controls throughout all stages—both positive controls (known interactions) and negative controls (non-transfected or irrelevant proteins)—to validate results effectively. Make sure to optimize each step based on specific requirements related to the proteins involved and desired outcomes from your study.

what cmo services do you offer for gmp plasmid dna manufacturing?
We offer gene and cell therapy manufacturing services, including GMP-grade plasmid DNA production. For more specific inquiries or to submit a request, please visit our website for detailed information on our offerings in plasmid manufacturing.

hi, do you sell default lentiviral samples with known titer for reference virus gostix to obtain gv readings?
The premade lentiviral samples available do not come with a precise titer; instead, they are labeled with a "greater than" value. Unfortunately, due to shipping challenges associated with lenti viruses, obtaining an exact control titer is not feasible. However, you can derive an initial estimate of infectious units (IFU) using the rough formula provided: - Each lentiviral particle (LP) contains about 2,000 molecules of p24. - One LP corresponds to approximately 8 x 10^-5 pg of p24. - One ng of p24 translates to roughly 1.25 x 10^7 LPs. - Typically, there is one IFU for every 100–1,000 LPs. This means that a supernatant titer around 10^7 IFU/ml would be approximately equal to between 10^9 and 10^10 LP/ml or about 80–800 ng p24/ml. After performing transductions with your first preparation, you can evaluate how close your calculations were and adjust accordingly for future preparations. This approach will help improve accuracy over time in estimating titers without needing a reference virus.

Why are the Tm values for gene-specific primers different from those of the vector portion overhang in my In-Fusion cloning?
The Tm (melting temperature) values for gene-specific primers and the vector portion overhang in In-Fusion cloning can differ due to several factors. Gene-specific primers are designed to bind specifically to the target insert sequence, which may have different nucleotide compositions or lengths compared to the vector's overlapping region. The presence of GC content, secondary structures, and specific sequences can all influence Tm values. Additionally, the overlapping regions with the vector are typically shorter than the full-length gene-specific primers, leading to differences in their thermal stability. This is why it's essential to consider these variations when designing your experiment and optimizing PCR conditions for successful cloning.

paav.cmv paav cmv plasmid kit available?
The pAAV.CMV plasmid kit is part of the AAV expression systems available, which includes various kits for creating AAV vectors. However, if you're looking for specific information about availability or particular components within those kits, it's best to check directly on the product website or contact customer support for detailed assistance.

how can I log account without password reset emails?
If you're unable to log into your account because you haven't received the password reset emails, there are a few steps you can take. First, check your spam or junk email folder in case the email was filtered there. Ensure that the email address associated with your account is entered correctly when requesting a reset. If you've confirmed that everything is correct and still haven't received an email, consider reaching out to customer support for further assistance. They may be able to help troubleshoot the issue or provide alternate options for accessing your account.