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

Daniela Nachmanson, Shenyi Lian, Elizabeth K. Schmidt, Michael J. Hipp, Kathryn T. Baker, Yuezheng Zhang, Maria Tretiakova, Kaitlyn Loubet-Senear, Brendan F. Kohrn, Jesse J. Salk, Scott R. Kennedy, Rosa Ana Risques
doi: https://doi.org/10.1101/207027
Daniela Nachmanson
1Department of Pathology, University of Washington, Seattle, WA 98195, USA
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Shenyi Lian
1Department of Pathology, University of Washington, Seattle, WA 98195, USA
2Department of Pathology, Key Laboratory of Carcinogenesis and Translational Research, Peking University Cancer Hospital and Institute, Beijing, PR, China
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Elizabeth K. Schmidt
1Department of Pathology, University of Washington, Seattle, WA 98195, USA
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Michael J. Hipp
1Department of Pathology, University of Washington, Seattle, WA 98195, USA
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Kathryn T. Baker
1Department of Pathology, University of Washington, Seattle, WA 98195, USA
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Yuezheng Zhang
1Department of Pathology, University of Washington, Seattle, WA 98195, USA
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Maria Tretiakova
1Department of Pathology, University of Washington, Seattle, WA 98195, USA
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Kaitlyn Loubet-Senear
1Department of Pathology, University of Washington, Seattle, WA 98195, USA
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Brendan F. Kohrn
1Department of Pathology, University of Washington, Seattle, WA 98195, USA
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Jesse J. Salk
3Department of Medicine, Division of Hematology and Oncology, University of Washington, Seattle, WA 98195, USA
4TwinStrand Biosciences, Seattle, WA 98121, USA
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Scott R. Kennedy
1Department of Pathology, University of Washington, Seattle, WA 98195, USA
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Rosa Ana Risques
1Department of Pathology, University of Washington, Seattle, WA 98195, USA
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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.

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The copyright holder for this preprint is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under a CC-BY 4.0 International license.
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Posted October 21, 2017.
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CRISPR-DS: an efficient, low DNA input method for ultra-accurate sequencing
Daniela Nachmanson, Shenyi Lian, Elizabeth K. Schmidt, Michael J. Hipp, Kathryn T. Baker, Yuezheng Zhang, Maria Tretiakova, Kaitlyn Loubet-Senear, Brendan F. Kohrn, Jesse J. Salk, Scott R. Kennedy, Rosa Ana Risques
bioRxiv 207027; doi: https://doi.org/10.1101/207027
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CRISPR-DS: an efficient, low DNA input method for ultra-accurate sequencing
Daniela Nachmanson, Shenyi Lian, Elizabeth K. Schmidt, Michael J. Hipp, Kathryn T. Baker, Yuezheng Zhang, Maria Tretiakova, Kaitlyn Loubet-Senear, Brendan F. Kohrn, Jesse J. Salk, Scott R. Kennedy, Rosa Ana Risques
bioRxiv 207027; doi: https://doi.org/10.1101/207027

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