Genotyping by next generation sequencing

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.