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Methods of Methylation detection not requiring bisulfite treatmetn - (May/24/2011 )

Hi Everyone,

I have very low yield DNA samples that I wish to perform methylation analysis on for a panel of genes. Given their low yield and the loss associated with bisulfite modification, does anybody know of any methylation detection methods not requiring bisulfite conversion? I was considering using a restriction enzyme based method. What have people's experiences been with these methods?



Your assistance is greatly appreciated.

-c.wright@uq.edu.au-

Restriction digestion using CpG methylation sensitive enzymes sounds straightforward, but the results usually is not that clear cut. You can also do ChIP using antibody against proteins that bind to methylated DNA such as MECP2 protein, or antibody that recognize 5m-cytosine.

-pcrman-

pcrman on Sun May 29 00:46:16 2011 said:


Restriction digestion using CpG methylation sensitive enzymes sounds straightforward, but the results usually is not that clear cut. You can also do ChIP using antibody against proteins that bind to methylated DNA such as MECP2 protein, or antibody that recognize 5m-cytosine.


Thank you for replying to my message. What are the major problems with restriction digestion? I want to validate results obtained using microarray analysis in body fluids of patients with resected cancer to see whether methylation can be detected in low yield samples. I unfortunately do not have access to all equipment required for antibody based methods. What else would you suggest?

-c.wright@uq.edu.au-