Combinatorial RNAi - double gene knockdown - (May/15/2008 )

Hi ,

Please tell me what to do -

I am trying to knockdown 2 proteins together - so i want to use 2 different RNAi's together at the same time - i have decided to split them in their concentrations and bring them upto 100 nm and am using the dharmafect reagent for transfection...and would be transfecting them twice . Can i increase their concentrations like use 100 nm of each at the same time is that possible...?????????
Has anyone tried something like this before..please let me know your comments or anything else i can do ???

I would do transfections individually just to be sure that bot siRNAs are active first, then do a co-transfection to test out if this will work for you. Not all cells receive/transfect siRNA in a single transfection experiment. The problem becomes more serious in a double transfection experiment. This will be further complicated with the degradation issue of the first siRNA. co-transfection gives the best chance for the same cell receive roughly the same amount of siRNA mixture, therefore it should be considered first.