Transfection of CRISPER Component into Cultured Cells and Drug Selection
2:31
Isolation of Clonal Populations
3:53
Screening Candidates by Dot Blot
6:30
Screening Candidates by Colony PCR
8:59
Results: Knockout Cell Lines Generated by Two CRISPER/Cas9 Methods
10:40
Conclusion
Transcript
The goal of this procedure is to isolate and identify clonal knockout cell lines generated by two types of CRIPSR/Cas9-mediated genome editing. One method detects protein knockout caused by nonhomologous end joining while the other detects genomic
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Recent advances in the ability to genetically manipulate somatic cell lines hold great potential for basic and applied research. Here, we present two approaches for CRISPR/Cas9 generated knockout production and screening in mammalian cell lines, with and without the use of selectable markers.