Keshet, on Apr 5 2009, 01:03 PM, said:
How long can I wait after seeing HeLa cells till transfection?
I heard that the optimal is 24 h, but I wander if it will affect my transfection if I will wait 72 …
I want to seed in 6-well plate before the weekend and to do the lipofectamine transfection in the beginning of the week.
Typically you can wait 5/6 hours after transfection before looking at your cells, as with most transfection reagents, maximum transfection is achieved after this time (using Jet Pei, Fugene6 etc). Obviously the longer you leave your cells, the more will be transfected, so it just depends on what you are going to do with them, and how many transfected cells you need.
With regards to longer transfections, 72 hours may be ok, the longer you leave it, the more chance of the cells losing the plasmid, as its not integrated into their DNA, in this case you could set up cells that stably express your gene of interest.
Also, i think you will be fine plating cells on the fri and transfecting on the mon, as long as the cells are not over-confluent, and healthy-looking, maybe try changing the transfection reagent before you do your transfection on the mon!
Hope this helps