What is the expected size range of fragmented, ds cDNA after library preparation with Nextera kits?

The Nextera kits from Illumina produce libraries with a size range of 300–1,000 bp. Please refer to the Nextera DNA Sample Preparation Guide or Nextera XT DNA Sample Preparation Guide for more specific details.Example Bioanalyzer electropherograms of cDNA libraries prepared for Illumina sequencing from 1 ng of Mouse Brain Total RNA (Panel A) or 1 ng of Human Universal Total RNA (Panel B) using a [...]

Do I have to scale down the Nextera XT DNA Sample Preparation Kit protocol when using 100–150 pg of ds cDNA?" "

No. Use 100–150 pg of ds cDNA generated with the SMARTer Ultra low kit in the input volume recommended in the Nextera XT DNA Sample Preparation Guide. Follow the rest of the protocol as written.General [...]

Can I use more than 150 pg of ds cDNA for the Nextera XT DNA Sample Preparation Kit?

In our hands, using 100–150 pg of input cDNA with the Nextera XT DNA Sample Preparation Kit generates DNA fragments with an optimal average size for Illumina cluster generation and sequencing. Using more than 150 pg of ds cDNA is not recommended since it generates significantly larger DNA fragments, which are suboptimal for Illumina cluster generation and sequencing.Example Bioanalyzer [...]

How do I pool cDNA libraries generated with the Low Input Library Prep kits for Illumina sequencing?

Follow the recommendations from Illumina for library pooling.General [...]

What is the expected size range of Covaris-sheared cDNA after library preparation?

cDNA generated with a SMARTer Ultra low kit that is sheared using Covaris technology and prepared with the Low Input Library Prep Kit typically has a size distribution of 150–600 bp with a peak at approximately 250–300 bp.Example Bioanalyzer electropherogram of a cDNA library prepared for Illumina sequencing. The cDNA library was prepared from 10 pg of Mouse Brain Total RNA using a SMARTer Ultra [...]