Haynes Lab:Notebook/HPK-CFP insertion into Gal4EED/Luc using CRISPR/2015/06/12

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Overview

Passaging HEK293 Gal4EED/luc line from PS-α to PS-β. Culturing in DMEM high glucose containing 10% FBS, 1% P/S, and 5 µg/mL puromycin.

For selection of gRNAs for CRISPR-mediated replacement, I'm going to go with g032, g034, g044 and g048 (René's gRNAs) in her vectors. If those don't work, will do genome engineering to construct gRNA oligos and insert into vectors based on the sequence of homology arms 1 and 2.