MSP question - (Sep/26/2005 )
Quick question. I had ordered MSP primers a while back before I decided to switch to BSP and just recently decided to try them. The problem that I am running into is that I don't think I used enough of my limited initial template (15 ng) and did not yield any bands on my methylated and non-methylated controls in one round of PCR.
As I no longer have more of the bisulfite treated samples is it experimentally sound to reamplify my first round products with the same msp primers? or would I likely run into artifact issues? I could always design the MSP to be off a nested 2 round PCR, but as I am just doing this out of curiosity I don't think it to be worth the effort.
Do you mean you want to reamplify your MSP or USP product with the same primer pair as the first round PCR?
I don't see anything wrong if the amplification is still in linear range.