methylation specific PCR (MSP) - reproducibilty? (Feb/04/2004 )
I know lot of people doing methylation research use MSP for methylation mapping. MSP depends on 1-2 bases in the primer to discriminate 5mC or non 5mC. I wonder how is reproducibilty of MSP since once I made a primer with a wrong base at the very 3' end and I could still got amplification of expected product. If you do 3 MSP, what is the chance of getting the same result?
than you for any input.
I once asked a similar question to people from the lab which invented the MSP technique and was told it works well at their hands. I think you have to tweak PCR conditions such as Ta, number of cycles. Unless you got them all optimized, this is a big concern.