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Chunk #49 — PROCEDURE — Co-transfection of CRISPR plasmids and HDR templates into HEK 293FT cells (optional) ● TIMING 3–4 d

Source
Genome engineering using the CRISPR-Cas9 system.
Embedded
yes

Text

medium warm.19Use option A in the table below for preparing the co-transfection of the HDR targeting plasmid with the Cas9 plasmid or option B for the co-transfection of ssODN with the Cas9 plasmid. To prepare transfection controls, see Step 9. If an sgRNA is cloned into pSpCas9(BB)-2A-GFP, cells may also be sorted by fluorescence. If you are using Cas9 nickase to mediate HDR, substitute pSpCas9(sgRNA) with pSpCas9n(sgRNA) from Step 5B(v).