Hi everyone!
SantaCruz's CRISPR/Cas9 KO plasmids contain GFP - has anyone tried selecting successful knockouts by FACS sorting? This would be as an alternative to additionally buying the HDR plasmid (which inserts the puromycin resistance gene into the genome) and selecting using puromycin.
Many thanks,
Sam
Simon Bornschein
Hey Sam,
I usually transfect/electroporate a gRNA-Cas9-GFP plasmid into cells and then sort on day 1 after transfection/electroporation for GFP+ cells. It depends a bit on the efficiency of the gRNA, but you can get full knockout population after the sort. So I assume it should also work for your Plasmid.
Best,
Simon
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