Improving sample quality for target enrichment and next-gen sequencing with the Agilent High Sensitivity DNA Kit and the Agilent SureSelect Target Enrichment Platform

Next-generation sequencing (NGS) has revolutionized the genetic landscape. It is a lengthy, labor-intensive process that yields results never before achieved. As a result, it is imperative that the quality of the DNA sample be evaluated from the start, as most NGS sample preparation protocols require PCR amplification to generate DNA libraries prior to sequencing. The likelihood of artifact generation could contribute to bias, affecting the potential results. The High Sensitivity DNA Kit used with the Agilent 2100 Bioanalyzer has been optimized with improved levels of detection. The improved sensitivity allows the numbers of library PCR cycles to be reduced, removing amplification bias and significantly improving the quality of NGS data with increased accuracy. This Application Note describes how the Agilent 2100 Bioanalyzer High Sensitivity DNA Kit can be used to provide quantitative and qualitative information about the DNA samples used in the Agilent SureSelect Target Enrichment System. of the sheared genomic DNA, and to assess the quality and size distribution of the PCR amplified sequencing library DNA. After post-hybridization amplification, the Agilent 2100 Bioanalyzer can be used to determine the quality and the concentration of the PCR-amplified capture DNA before sequencing. This Application Note describes how the High Sensitivity DNA kit and the Agilent 2100 Bioanalyzer can be used before sequencing to reduce the number of required PCR cycles. This reduces amplification bias, thus improving the quality of DNA libraries created during the SureSelect Target Enrichment workflow.