siRNA + DNA transfection - (Apr/25/2007 )

Hi!

I need to carry out a luciferase assay in previuously siRNA-transfected cells. First, I'd have to transfect cells with siRNA (using Lipofectamine 2000) and then I would have to transfect luciferase and renile constructs (using Lipofectamine + Plus Reagent). That is what works best in my experimental conditions.... My question is: Are my cells going to suffer a "big-bang" owing to so much transfection???? What could I do?

Thank you for your help!

Hi yarince,

The proposed transfection should not be a problem to many commonly used cells. I have done simutaneouse siRNA and DNA transfection using lipofectamine and staggering transfections as well but in the reverse order of yours. I first transfected DNA using lipofectamine + plus, the 2nd day, I passed the cells and did siRNA transfection using lipofectamine 2k. The cells tolearated well the transfection. Cells I used are cancer cell lines.

Hope that helps.

Thank you pcrman!!!

I'll try next week your advices...