siRNA transfection: DNA conc optimization problem - (May/14/2008 )
im using Cho-K cell line for transfection studies. previously i did optimized DNA conc. which was 50ng (for RT-PCR) using Lipofectamine 2000. but now when im using siRNA for knockdown analysis. im getting nothing, would anybody tell me where im wrong or what could be the possible reasons for not getting RT expression????????????????
I am confused. DNA template for RT-PCR or for transfection. RT expression??
If your question is related to transfection, you have to tell us what size of the multiwell plate you are using. 50 ng is not a lot of DNA. You may need to get to 0.1-0.3 ug range for a 96 well plate with Lipo 2000.