CRISPR-DS: An efficient, low DNA input method for ultra-accurate sequencing

Abstract

Conventional hybrid-capture based Next-Generation Sequencing suffers from frequent errors, PCR bias due to variable fragment length, and inefficient target enrichment. To address these shortcomings, we combined ultra-accurate Duplex Sequencing with CRISPR/Cas9 excision of target regions, which allows enrichment by size selection prior to library preparation. This high efficiency method, which we term CRISPR-DS, improves targeted sequence recovery, reduces PCR bias, and maximizes read usability from minimal DNA inputs.