RT Journal Article
SR Electronic
T1 CODEC enables ‘single duplex’ sequencing
JF bioRxiv
FD Cold Spring Harbor Laboratory
SP 2021.06.11.448110
DO 10.1101/2021.06.11.448110
A1 Bae, Jin H.
A1 Liu, Ruolin
A1 Nguyen, Erica
A1 Rhoades, Justin
A1 Blewett, Timothy
A1 Xiong, Kan
A1 Shea, Douglas
A1 Gydush, Gregory
A1 Tabrizi, Shervin
A1 An, Zhenyi
A1 Patel, Sahil
A1 Makrigiorgos, G. Mike
A1 Golub, Todd R.
A1 Adalsteinsson, Viktor A.
YR 2021
UL http://biorxiv.org/content/early/2021/06/12/2021.06.11.448110.abstract
AB Detecting mutations as rare as a single molecule is crucial in many fields such as cancer diagnostics and aging research but remains challenging. Third generation sequencers can read a double-stranded DNA molecule (a ‘single duplex’) in whole to identify true mutations on both strands apart from false mutations on either strand but with limited accuracy and throughput. Although next generation sequencing (NGS) can track dissociated strands with Duplex Sequencing, the need to sequence each strand independently severely diminishes its throughput. Here, we developed a hybrid method called Concatenating Original Duplex for Error Correction (CODEC) that combines the massively parallel nature of NGS with the single-molecule capability of third generation sequencing. CODEC physically links both strands to enable NGS to sequence a single duplex with a single read pair. By comparing CODEC and Duplex Sequencing, we showed that CODEC achieved a similar error rate (10−6) with 100 times fewer reads and conferred ‘single duplex’ resolution to most major NGS workflows.Competing Interest StatementThe authors have filed a patent application on this method. V.A.A. is a member of the scientific advisory boards of AGCT GmbH and Bertis Inc. T.R.G. has advisor roles at Foundation Medicine, GlaxoSmithKline, and Sherlock Biosciences.