Lipofectamine and transfection - (Jul/12/2011 )

I did a transfection experiment yesterday using Lipofectamine 2000. Unfortunately I waited about 35-40 minutes (instead of the recommended 5-30 minutes) for my Lipofectamine and Optimem master mix to incubate at room temp before I added my DNA. Invitrogen said the protein expression may be reduced, but I'd like to know how much reduced--that is, if I can still expect decent protein expression, if at all. I used GFP promoter to express F1L mutants (which are about 50 kda total), and I expressed them in HEK 293 T cells at 80-90% confluency. Thanks!

You should be fine. The encapsulated DNA should be pretty stable even at room temperature.

pcrman on Wed Jul 13 03:56:52 2011 said:

yeah, and 293T are very easy to transfect, so you should have no problem at all.