This document describes how the Agilent High Sensitivity DNA Kit and the Agilent SureSelect Target Enrichment Platform can improve sample quality for target enrichment and next-generation sequencing. By utilizing improved detection sensitivity, the number of library PCR cycles can be reduced, eliminating amplification bias and significantly improving the quality and accuracy of NGS data.
This document is an application note detailing the use of the Agilent High Sensitivity DNA Kit in conjunction with the Agilent 2100 Bioanalyzer. It focuses on improving sample quality for target enrichment and next-generation sequencing (NGS) workflows, specifically the SureSelect Target Enrichment Platform.
The note explains that the High Sensitivity DNA Kit allows for the detection and quantification of DNA libraries with as few as four PCR cycles. This capability helps reduce amplification bias and artifacts compared to higher cycle counts, thereby improving the accuracy of NGS data. The document presents experimental results comparing the kit's performance against fluorometer-based quantification methods.
The High Sensitivity DNA Kit offers improved sensitivity for checking sample size and quantity down to a concentration of 100 pg/L.
Above 14 PCR cycles, the increase in DNA concentration is no longer linear and becomes saturated.
An integration region from 100 to 2000 bp is used for all samples for smear quantification.