Transient transfection of MEFs - (Apr/30/2007 )
Hi, my MEF cells are immortalized by SV40 transformation. I am trying to do transient transfection, I used Fugene 6, which gives me a poor efficiency. Anyone has good suggestion? I hope to get a efficiency close to 50%.
Thanks a lot.
-helpme7-
Have you optimized the transfection condition with regard to 1) DNA/lipid ratios, 2) cell density, and 3) DNA dosages? Other options will be LipofectAMINE 2000 (Invitrogen), transfectam (Promega). However, you still need to do an optimization on these reagents.
-genehunter-1-
QUOTE (genehunter-1 @ Apr 30 2007, 09:27 AM)
Have you optimized the transfection condition with regard to 1) DNA/lipid ratios, 2) cell density, and 3) DNA dosages? Other options will be LipofectAMINE 2000 (Invitrogen), transfectam (Promega). However, you still need to do an optimization on these reagents.
Thanks, I think do try to optimize a few conditions, not extensively, but the same low efficiency. Some one said MEFs are diverse from each other, for the one I got they never transient transfect more than 15%. I am think making retrovirus now.
-helpme7-
QUOTE (helpme7 @ Apr 30 2007, 08:42 AM)
QUOTE (genehunter-1 @ Apr 30 2007, 09:27 AM)
Have you optimized the transfection condition with regard to 1) DNA/lipid ratios, 2) cell density, and 3) DNA dosages? Other options will be LipofectAMINE 2000 (Invitrogen), transfectam (Promega). However, you still need to do an optimization on these reagents.
Thanks, I think do try to optimize a few conditions, not extensively, but the same low efficiency. Some one said MEFs are diverse from each other, for the one I got they never transient transfect more than 15%. I am think making retrovirus now.
I obtained good results by using DreamFect Gold or DramFect Gold + CombiMag (OZBiosciences) as transfection reagents. I got around 50% transfection efficiency in MEFs.
-cec44-