suggested siRNA concentrations - (Oct/23/2008 )

Hallo out there!

I am working with siRNA and do double transfections with plasmid DNA in HeLa cells.

I have decided which concentrations of DNA plasmid work better, although when i transfect in parallel with siRNA, the efficiency of siRNA transfection is higher than the plasmid's efficiency in my hands!

For the siRNA i use 40pmol for 1 well of 12-well plate. The transfections I do with Lipofectamine.
I believe that this is rather a big amount and that the effect could also happen with lesser.

I know from the literature that the lesser amounts of siRNA you use, the better for diminishing off-target effects.

So has anyone an idea, from how much should I begin and test?

Thank you in advance

40 pmol (40 nM/L) is in the normal range and should be fine. siRNA works quite well from 10-100 nM/L