hello u all!!
I ve just got my result from the MSP. the universal methyated DNA is indid methylated (!!) but a much weaker band is also seen from the unmethylated primers?
does it mean that the primers are not specific in those annealing tempratures?
what it the best way to continue?
universal meth control is unmethylated?
1 reply to this topic
Posted 15 March 2009 - 09:55 AM
It is not a surprise that you got amplification from methylated control DNA by unmethylated primers because the primers rely on a few nucleotides to differentiate unmethylated from methylated DNA. You can try raising the annealing temperature a little bit or cutting the cycle number down by a few cycles.