principle of transfection - wants to know the optimal time needed for transfection (Jan/05/2006 )
i have transfected HeLa cells using Lipo2000.
After transfecting with different constructs, I had chnaged the medium (DMEM+FCS)
after 6 hrs to complete medium.
My question is 4-6 hrs sufficient time for a DNA complex to enter the cells?
Since due to toxicity issue, i had chnaged the medium, and now I am scared, whether transfection will be a success or not ?
The cells were nicely adhered to the surface, after a day does that mean my transfection was successful?
Is there any other way to know about successful transfection?
thanks for the help.
This should be a good enought time period for the liposomes to have entered the cells. I usually let the transfection go for 5-7 hours before changing the media using Lipo2000 as well.
I usually go down to about 3-4 hrs, which is plenty. I believe it helps my cells survive, because of decreased exposure to toxic agents (which transfection agents are to some degree) and also to have them sooner back in complete medium with serum.