What are the advantages of library prep with the ThruPLEX DNA-Seq Kit?

The ThruPLEX DNA-Seq Kits (Cat. # R400674–R400677) have the following advanced features: High performance from low inputs—generate libraries from 50 pg to 50 ng of input material Fast and simple workflow—three-step, single-tube or single-well protocol takes only about two hours, no transfers necessary Compatible with major target enrichment platforms—suitable for use with Agilent SureSelect, [...]

How should I set up the Peak Incident Power (W) for the Covaris S220 system?

The intensity for the S220 Covaris protocol is an equivalent to Peak Incident Power (W) set at 175. General [...]

What type of Covaris machine did you use to optimize the shearing parameters?

Covaris shearing parameters, provided in the user manuals of the SMARTer Ultra low kits, were optimized using a Covaris S220 Focused-ultrasonicator. If you are using another type of Covaris apparatus, please consult the manufacturers for the recommended parameters to ensure DNA is in the size range of 100–500 bp with a peak at approximately 200 bp.General [...]

What is the expected size range of Covaris-sheared double-stranded (ds) cDNA?

Covaris-sheared ds cDNA should span 100–500 bp with a peak of approximately 200 bp. To ensure optimal Covaris ds cDNA shearing: Do not load more than 75 µl of ds cDNA per 100 µl Covaris tube. Avoid introducing air bubbles when loading the ds cDNA in the Covaris tube. Example Bioanalyzer electropherogram of Covaris-sheared ds cDNA. Recommendations for Covaris DNA shearing conditions can be [...]

What method should I use to prepare cDNA generated with SMARTer Ultra low kits for sequencing?

We recommend two preparation methods for Illumina sequencing platforms: Covaris shearing followed by library construction with the ThruPLEX DNA-Seq Kit (Cat. # R400674–R400677). This kit is compatible with 50 pg–50 ng of fragmented, double-stranded DNA (<1,000 bp), allows multiplexing, and has been validated for downstream Illumina sequencing platforms. The Nextera® XT DNA Sample Preparation [...]