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?
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.
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!
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.
Answered 1 year ago
Dr. SD Udayappan
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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.
Answered 1 year ago
Dr. SD Udayappan
2 Comments
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!
Answered 11 months ago
Cornelis Eriksson
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.
Answered 1 year ago
Dr. SD Udayappan
Can you help?