RT Journal Article
SR Electronic
T1 Targeted genome fragmentation with CRISPR/Cas9 improves hybridization capture, reduces PCR bias, and enables efficient high-accuracy sequencing of small targets
JF bioRxiv
FD Cold Spring Harbor Laboratory
SP 207027
DO 10.1101/207027
A1 Daniela Nachmanson
A1 Shenyi Lian
A1 Elizabeth K. Schmidt
A1 Michael J. Hipp
A1 Kathryn T. Baker
A1 Yuezheng Zhang
A1 Maria Tretiakova
A1 Kaitlyn Loubet-Senear
A1 Brendan F. Kohrn
A1 Jesse J. Salk
A1 Scott R. Kennedy
A1 Rosa Ana Risques
YR 2018
UL http://biorxiv.org/content/early/2018/01/12/207027.1.abstract
AB Current next-generation sequencing techniques suffer from inefficient target enrichment and frequent errors. To address these issues, we have developed a targeted genome fragmentation approach based on CRISPR/Cas9 digestion. By designing all fragments to similar lengths, regions of interest can be size-selected prior to library preparation, increasing hybridization capture efficiency. Additionally, homogenous length fragments reduce PCR bias and maximize read usability. We combine this novel target enrichment approach with ultra-accurate Duplex Sequencing. The result, termed CRISPR-DS, is a robust targeted sequencing technique that overcomes the inherent challenges of small target enrichment and enables the detection of ultra-low frequency mutations with small DNA inputs.DSDuplex SequencingDCSDouble-stranded consensus sequenceSSCSSingle-stranded consensus sequencegRNAGuide RNAcrRNACRISPR RNAtracrRNATrans-activating crRNANGSNext-generation SequencingngNanogrambpBasepairssDNASingle-stranded DNAdsDNADouble-stranded DNADINDNA integrity number