Chunk #53 — 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

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▲ CRITICAL STEP Cells are very fragile at this stage, and harsh pipetting can cause cell death. 28Incubate the mixture for 24 h. At this point, transfection efficiency can be estimated from the fraction of fluorescent cells in the positive transfection control. Nucleofection typically results in >70–80% transfection efficiency.