Methylation Specific PCR Protocol

[joejeo]

Anyone know how to do Methylation Specific PCR Protocol?

thanks in advance

[Da_vinci]

I found a great Methylation PCR protocol here:

[Only registered users see links. ]

[lindapiekuse]

IS someone doing DMSO methylation specific PCR? Just really a lot of trouble of it.
Thankx

[vladimirec]

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

[hermawan]

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

[molpro]

actually i have also done methylation PCr.
the best is to digest the template with EcoRI and further proceed for the pcr.

[Chromatin-girl]

By template you mean bisulfite converted DNA?