Stable siRNA transfection

Labnovice · #1 (**permalink**) 04-20-2008, 10:57 PM

HI,

I do a lot of siRNA transfection for expts like PI staining, protein extracts, western blot etc wherein I require a lot of cells.
The main disadvantage is that I have to do all these expts post 48 hours due to the transient nature of siRNA knockdown and also use up a lot of the expnsive Lipofectamine 2000 every week.

Is there any way to stably knockdown my gene of interest in the stable ovarian cancer cell line I use, using siRNAs? Does it take a lot of time?
Please suggest with a little detail coz I cant grasp all the promoter, construct language very quickly

Also can anyone recommend any other transfection reagents they use for siRNA treatment which is good and relatively cheaper?

Thanks,

biology · #2 (**permalink**) 05-22-2009, 12:34 PM

Hi,

I have to start the sirna transfection experiments. Can anyone help me with what are the precautions i have to follow in these experiments, as RNAses are ubiquitously present.
I have some basic knowledge of transfection and created stable cell lines also.

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Stable siRNA transfection

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Stable siRNA transfection - Discuss and Post Questions in this forum about RNA interference and siRNA. RNAi and siRNA transfection, inhibition of endogenous genes, and miRNA and shRNA.