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

Nachmanson, D.; Lian, S.; Schmidt, E. K.; Hipp, M. J.; Baker, K. T.; Zhang, Y.; Tretiakova, M.; Loubet-Senear, K.; Kohrn, B. F.; Salk, J. J.; Kennedy, S. R.; Risques, R. A.

doi:10.1101/207027

bioRxiv · 10.1101/207027

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

Nachmanson, D.·Lian, S.·Schmidt, E. K.·Hipp, M. J.·Baker, K. T.·Zhang, Y.·Tretiakova, M.·Loubet-Senear, K.·Kohrn, B. F.·Salk, J. J.·Kennedy, S. R.·Risques, R. A.

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Abstract

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.

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Nachmanson, D., Lian, S., Schmidt, E. K., Hipp, M. J., Baker, K. T., Zhang, Y., Tretiakova, M., Loubet-Senear, K., Kohrn, B. F., Salk, J. J., Kennedy, S. R., Risques, R. A.. 2017-10-21. CRISPR-DS: An efficient, low DNA input method for ultra-accurate sequencing. https://doi.org/10.1101/207027

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