paperKB
coga / coga-kb
Processing
Help
Sign in

Chunk #58 — PROCEDURE — hESC (HUES 9) culture and transfection ● TIMING 3–4 d

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

Text

? TROUBLESHOOTING 51At 48–72 h post transfection, dissociate the cells with Accutase and resuspend them gently in a 5× volume of mTeSR1. Reserve a fraction of the resuspended cells at this stage for replating (Steps 41 and 42; make sure to add ROCK inhibitor for each passaging), for downstream applications or clonal isolation (Steps 54–70), and use the remaining cells for genotyping (Steps 71–126).