HepG2 siRNA transfection - (Aug/07/2009 )

Hello!

Im trying to transfect HepG2 cells with siRNA. First i tried with Lipofectamine 2000 and siRNA 50nM, but the lipofectamine was affecting the protein im studying. Now im trying to perform the transfection with Nucleofector. Following the Amaxa protocol and using 1x10^6 cells per sample and the high viability protocol, most of my cells are dying. They do not reccomend to use more cells per sample, because it would affect the transfection efficiency. Im using medium with Pen/Strep, but i think its not toxic in this case. Have anyone tried to do the transfection with a higher number of cells or succed with HepG2 and siRNA transfection using Nucleofector?

Thanks for your help!

Itziar

why kill your cells with electroporation? how much cells would you like to transfect? Try a cationic lipid like saint and optimize your protocol. I think it is easier?