Abstract 3564: A new method for preparation of low-input, PCR-free next generation sequencing libraries

Sequence analysis of PCR-free Next Generation Sequencing (NGS) libraries demonstrates improved evenness of coverage relative to NGS libraries that have undergone PCR amplification. However, existing PCR-free methods require 1ug or more of input DNA quantity in order to obtain sufficient library yield which restricts such analysis when DNA sample quantity is insufficient. To expand the applicability of PCR-free sequencing, Swift Biosciences has developed a novel adapter attachment chemistry that is more efficient than existing methods. This new technology, Accel-NGS™, results in libraries that capture a higher complexity of each sample which enables a lower input DNA requirement. For Illumina systems, PCR-free Accel-NGS libraries can be prepared from 100ng of input DNA for direct sequencing. PCR-free Accel-NGS libraries demonstrate even coverage across a broad GC content as well as when analyzing whole genome sequence coverage. To enable library preparation from even less DNA, Accel-NGS libraries can also be prepared with a low number of PCR cycles for direct sequencing. The Accel-NGS method with PCR results in more even coverage and less PCR duplicates when compared to existing methods that use PCR. For exome hybridization capture, comparable coverage was achieved for Accel-NGS libraries from 100ng DNA input when compared to an existing method that requires 1ug of DNA input. The Accel-NGS method is fast and automatable; additional advantages when applied to clinical samples are under investigation. PCR-free Accel-NGS libraries can also be prepared for Ion Torrent systems from 5ng of input DNA with comparable sequence quality to existing methods. Citation Format: Laurie Kurihara, L. Banks, S. Chupreta, C. Couture, V. Kelchner, J.Laliberte, S. Sandhu, R. Spurbeck, V. Makarov. A new method for preparation of low-input, PCR-free next generation sequencing libraries. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 3564. doi:10.1158/1538-7445.AM2014-3564