Can I use more than 150 pg of ds cDNA for the Nextera XT DNA Sample Preparation Kit?
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 electropherograms of RNA-seq libraries generated from either 130 pg (Panel A) or 1 ng (Panel B) of cDNA generated using a SMARTer Ultra low kit. 130 pg of input cDNA generates libraries of optimal size for Illumina cluster generation and sequencing.