What are the benefits and application of two-step RT-qPCR?

Two-step RT-qPCR allows for the preparation of total cDNA by reverse transcription using universal primers that recognize all mRNA molecules, such as random hexamers or oligo-dT primers. The resulting cDNA can be used for the detection of a variety of transcripts. When there is an overabundance of total RNA in the reaction, oligo-dT primers are recommended for two-step RT-qPCR rather than [...]

What are the benefits and application of one-step RT-qPCR?

One-step RT-qPCR uses gene-specific primers for reverse transcription and allows highly sensitive detection of a specific gene. When there is an overabundance of total RNA in the reaction, one-step RT-qPCR van provide highly efficient amplification even in the presence of large amounts of total RNA. One-step RT-qPCR is usually used to analyze a single gene. For high-throughput analyses of many [...]

Can I purchase RCU buffer from 740910 NucleoSpin RNA Clean-up Maxi?

No. However, you can buy bufferLysis Buffer RA1, which is comparable to buffer RCU. Also it is the same volume as in the NucleoSpin RNA Clean up Maxi kit . You can proceed with buffer RA1 just like with the RCU concentrate and dilute it with 180 mL ethanol. It is not recommended to use more ethanol to dilute the buffer. The kit is tested and optimized to show best performance. Changing the [...]

Do you have any special recommendations for isolating viral RNA from breast milk?

Both milk and plasma are protein-rich fluids and can be treated as similar/equivalent biological fluids with regard to their biochemical properties. Therefore we recommend the NucleoSpin RNA Virus Kit (REF 740956/.50/.250) for this application. The NucleoSpin RNA Virus kit is designed for highly pure viral nucleic acids from fluid biological samples, for example plasma or serum. Please note that [...]

Is the RLT buffer from Qiagen compatible with NucleoMag Virus? Customer is isolating Covid-19 virus from various samples stored in the RLT buffer.

Yes. The RLT can be used in combination with NucleoMag RNA: * Clear lysate: centrifuge at 5,600 x g for 5 min, transfer supernatant to a new tube. * Add 1 vol of 70% EtOH to supernatant, as per Qiagen protocol. * Add 28 µL B-Beads per 350 µl sample * Mix well (e.g shaking for 5 min) * Put on magnetic stand for 2 min. Remove supernatant * Dry beads for 5 min. * Proceed with step 4 of the [...]