Hanqin Li, Oriol Busquets, Yogendra Verma, Khaja Mohieddin Syed, Nitzan Kutnowski, Gabriella R Pangilinan, Luke A Gilbert, Helen S Bateup, Donald C Rio, Dirk Hockemeyer, Frank Soldner (2022) Highly efficient generation of isogenic pluripotent stem cell models using prime editing eLife 11:e79208
License: This is an open access collection distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited
Protocol status: Working
We use this collection and it's working
Created: February 03, 2022
Last Modified: May 31, 2024
Collection Integer ID: 57787
Keywords: ASAPCRN, genotyping procedure, next generation sequencing, collection overview preparing of genomic dna, target genomic region by pcr preparing sample, target genomic region, hpscs culture, target genomic region by pcr, cell lysate from fac, preparing crude cell lysate, genomic dna, term hpsc, hpsc, crude cell lysate, pcr preparing sample, preparing cell, cultured cell, preparing crude cell, hipsc, ngs crispresso analysis, preparing sample, described procedure, hesc, crispresso analysis
Funders Acknowledgements:
Aligning Science Across Parkinson's
Grant ID: ASAP-000486
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.
