mRNA expression low/no effect of the siRNA - (Mar/29/2007 )
I need any advices. I check the effect of siRNA using two-step real-time PCR but the mRNA expression of my gene is very low = 30 cycle but GAPDH = 18 cycle. And maybe I have difficulty in seeing the effect of the siRNA (I have checked 5 different siRNAs). I use FastLane cDNA (cDNA directly from cultured cells without RNA purification for use in two-step RT-PCR). What should I do?
I wil be grateful for help.
Is anyone in your lab using this metod? And if yes - what is the efficiency?
Does siRNA are designed by you or by company?
Maybie you should try extract
RNA with TRIreagent from Roche and make normal RT...
and mRNA of what gene are you silencing?