Haynes Lab:Notebook/HPK-CFP insertion into Gal4EED/Luc using CRISPR/2015/07/13
|Transfection of Gal4EED-Luc HEK293||Main project page|
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Going to perform four transfections of Gal4EED-Luc cell line HEK293 in 6-well plates, using four combinations of donor plasmid and gRNAs/Cas9 plasmids.
Calculations for making plasmid solutions:
I'll be performing transfections using the Lipofectamine LTX kit, following their instructions with one addition: centrifugation of the plate immediately post-transfection, 30 min @ 126 g and 37 °C. Plate centrifugation has been shown to greatly enhance transfection efficiency in other cell lines.