Making a GPCR Stable - DNA amount? - (Jun/16/2006 )
I'm trying to make a stable in CHO-K1 cells of a GPCR. I am using Lipofectamine 2000 to transfect.
Was wondering if anyone out there tried making stables of GPCRs and what amounts of DNA and Lipofectamine they used which gave successul stable clones?
The manufacturer's protocol says 4ug of DNA for a 6 well plate which I am thinking sounds high?
For transfection using lipofectamine 2000, I use 2ug DNA per well (6 well plate). Usually get more than 90% transfected.
Do you use 2 ug DNA with 3 ul Lipofectamine 2000?