• 云顶集团3118(中国)

    Genotyping by next generation sequencing

    Yogendra Verma, Hanqin Li, Dirk Hockemeyer, Frank Soldner

    Published: 2022-09-06 DOI: 10.17504/protocols.io.b4n3qvgn
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    Abstract

    This collection describes a standard genotyping procedure using next generation sequencing (NGS) in the Hockemeyer lab.

    Collection overview

    Preparing of genomic DNA from in vitro cultured cells

    A. Preparing crude cell lysate directly from hPSCs culture
    
    B. Preparing crude cell lysate from dissociated cells
    
    C. Preparing cell lysate from FACS-sorted cells
    

    Amplifying the target genomic region by PCR

    Preparing samples for NGS

    CRISPResso analysis

    General notes:

    1. Throughout these protocols, the term hPSC is used to collectively refer to both hiPSCs and hESCs. All described procedures have been tested and work equally well for hiPSCs and hESCs.

    Steps

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