I'm a beginner in RNA isolation and reverse transcription. We are planning use these methods to evaluate siRNA transfection. I isolated RNA and did reverse transcription (Invitrogen kit). I made triplicates from each RNA sample. After reverse transcription I did simple PCR, and I supposed that PCR products of cDNA from one RNA samples should have approx. same strong bands, bud I had even 5-fold differences within triplicates on agarose gels.
So, do you think I did something wrong during procedures or whole idea is wrong?
Thanks answers and please excuse my English.
Edited by majihec, 12 November 2010 - 04:29 AM.