siRNA transfection with lipofectamine 2000 - (May/05/2011 )
I've been working with siRNAs just for a couple of weeks and i have more doubts that answers! When i transfected siRNA with lipofectamine with the amounts that appear in the protocol, all my cells died. So i asked one of my lab mates and he told me to use just 1ul lipofectamine for 20pmol of siRNA (that's the protocol specifications)but instead for each well, use that amount for three wells.
Now i have to do the same kind of transfection but in a 6 well plate and i wonder if you guys know which would be the best amount to use, because the protocol tell me to use 100pmol of siRNA and 5ul of lipofectamine, and i think that,maybe, would be a little high...
I hope you can help me.
Thank you very much
ps: I love this website by the way!!
There is a safe and workable range for the amount of any transfection reagent. If toxicity is a problem, you can use the lowest suggested amount and also seed the cells at a higher density. You can also try less toxic reagents such as RNAimax from invitrogen and HiPerFect from Qiagen.