How “strong” is the PCR product of methylation specific PCR?

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5 years ago

Daniel McKenzie

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How “strong” is the PCR product of methylation specific PCR?

Does anyone know how “strong” the PCR product of methylation specific PCR is? I kept my PCR products at 4C for about 3 weeks and then at room temperature for another week. Will I be able to use them for sequencing?

Experiment: PCR Methylation specific PCR - Mammalian DNA

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**Dr. SD Udayappan** · Answer 1 · 2019-06-26T23:41:09+00:00

PCR products are incredibly stable. So, it should work for sequencing. Moreover, the newly ampiflied DNA won't get methylated in a PCR reaction as there is no methylating enzyme and polymerases don't methylate. The only thing you should take into account is; multiple freeze-thaw cycles (which can cause shearing). Hope this will help you.

**Cornelis Eriksson** · Answer 2 · 2020-05-11T09:30:53+00:00

Nope, DNAses are everywhere and at room temperature they will be active. The DNA in your sample will most likely be all cleaved up. It may be a neat experiment to amplify a gene like rp49 to see what you have but that may be a waste of expensive reagents and time!

**Dr. SD Udayappan** · Answer 3 · 2019-06-26T23:41:09+00:00

PCR products are incredibly stable. So, it should work for sequencing. Moreover, the newly ampiflied DNA won't get methylated in a PCR reaction as there is no methylating enzyme and polymerases don't methylate. The only thing you should take into account is; multiple freeze-thaw cycles (which can cause shearing). Hope this will help you.