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universal meth control is unmethylated?

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#1 kerenshi



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Posted 15 March 2009 - 03:28 AM

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?
k :D

#2 pcrman



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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.

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