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Methylation & PCR - (Oct/07/2005 )

I need to design a few RFLP assays, and I've never done this sort of thing before. The restriction enzyme I want to use is completely blocked by CpG methylation.

However, I'm going to be digesting PCR product. PCR amplicons aren't methylated, right? If I remember, one needs something like methyl transferase in order to have nucleotides methylated.

So, that means, that the restriction enzyme should work fine on PCR product, right?


You're absolutely right. After PCR, cytosines with methyl group lose their methylation.