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Epigenetics Forum: DNA Methylation, Histone and Chromatin Study An Epigenetics Forum Dealing with DNA Methylation, the study of histones and chromatin


Methylation Specific PCR Protocol

Methylation Specific PCR Protocol - Epigenetics Forum: DNA Methylation, Histone and Chromatin Study

Methylation Specific PCR Protocol - An Epigenetics Forum Dealing with DNA Methylation, the study of histones and chromatin


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  #1  
Old 06-20-2006, 08:59 AM
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Question Methylation Specific PCR Protocol



Anyone know how to do Methylation Specific PCR Protocol?

thanks in advance
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Old 07-07-2006, 08:07 PM
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Default Re: Methylation Specific PCR Protocol

I found a great Methylation PCR protocol here:

[Only registered users see links. ]
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Old 01-19-2010, 06:40 PM
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Default Re: Methylation Specific PCR Protocol

IS someone doing DMSO methylation specific PCR? Just really a lot of trouble of it.
Thankx
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Old 03-19-2010, 04:42 PM
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Default Re: Methylation Specific PCR Protocol

Could someone give little more detailed protocol??
Ex. How much DNA, how much Sodium bisulfate, how long is the treatment. ....

I just need to know how much DNA can I convert with Sodium bisulfate at once.

thanks

v
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Old 05-22-2010, 08:09 PM
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Default Re: Methylation Specific PCR Protocol

Dear All,

I'm hermawan, I am a new member here. I'm doing a research dealing with MSP.
I have 2 primer for methylated and 1 primer for unmethylated.
I'm still doing optimization for MSP. For this optimization, I use DNA template for methylated and unmethylated using Methylated and Unmethylated DNA control from Qiagen respectively.
I got a good result (clear band) in unmethylated primer. For methylated primer, I got a band in the right place (in accordance with its amplicon size), but the intensity of brightness is still less than unmethylated primer, it is still faint. I don't know why.
For this MSP, I have done temperature gradient PCR to choose the best annealling temperature based on Tm Forward and reverse primer given by company where I order those primer (eurogentec ait, singapore)
Please give me any suggestion, what should I do next to get a better result in MSP for my methylated primer?
Thank you
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Old 05-27-2010, 10:45 AM
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Default Re: Methylation Specific PCR Protocol

actually i have also done methylation PCr.
the best is to digest the template with EcoRI and further proceed for the pcr.
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Old 07-01-2010, 06:18 PM
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Default Re: Methylation Specific PCR Protocol

By template you mean bisulfite converted DNA?
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