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


siRNA optimization

siRNA optimization - RNAi and SiRNA Forum

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


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Old 10-01-2008, 09:55 AM
Pipette Filler
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Question siRNA optimization



Hi,

I am starting a experiment where i wish to use siRNA to target a gene in MCF-7 cells.

I have a pre-validated siRNA and a negative scrambled control from Ambion.

I began my optimization by using a range of siRNA:transfection reagent ratios and anaylzed the gene expression using semi-quantitative RT-PCR. However, i have now noticed that alot of people seem to use real-time and i was wondering if my optimization results will be accurate enough to allow me to proceed with the next stages of the experiment or should i do a real-time pcr to confirm?

Also i was just wondering if its necessary to have a positive control as i don't want to do all the work & then be told that i should have had one!

Many thanks in advance!
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