Abstract 4151: A novel, fast, and efficient technology for the preparation of high quality NGS libraries.

Abstract

Abstract Today's next-generation sequencing (NGS) investigators and clinicians must extract and analyze complex sequence data from a wide range of sample types. Preparing fragment libraries from these samples for whole genome, exome or targeted gene panel sequencing by NGS is a laborious process, characterized by long prep times and sample compatibility constraints due to DNA input quantity, genomic complexity, and molecular integrity. The workflows for most commercially-available kits are variations on traditional modifications for the repair, adaptation, and selection of DNA fragments, requiring 2-4 hours of preparation time before sequencing. As a consequence, NGS users cite sample preparation as the second most time-consuming step and bottleneck in their NGS workflows (following data analysis and interpretation; Bioinformatics, Oct. 2011). We present a novel technology that enables faster and more efficient NGS fragment library preparation, which increases yield and is compatible with many sample types, including damaged, cross-linked samples derived from FFPE or ChIP or samples with limiting DNA quantity. This method conveys these advantages because it does not require intact, double-stranded DNA for adapter ligation, and the streamlined workflow results in significant time savings relative to the workflows of commercial products. Side-by-side comparisons of sequencing data from libraries prepared using this new technology versus those from existing commercial products demonstrates equivalent or improved genome coverage, and suitability for damaged or cross-linked samples. Citation Format: Laurie Kurihara, Ron Beaubien, Jeff Perry, Vanessa Kelchner, Drew McUsic, Anjali Mishra, Julie Laliberte, Sergey Chupreta, Vladimir Makarov. A novel, fast, and efficient technology for the preparation of high quality NGS libraries. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 4151. doi:10.1158/1538-7445.AM2013-4151