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How to qualitatively measure DNA methylation of gene promoter

DNA methylation Gene Promoter

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#1 sultan

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Posted 22 January 2014 - 05:10 AM

Hi 

Can any body help me. Weather how can I detect particular gene promoter is methylated or not only quantitatively.

Cheers 



#2 Wunder

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Posted 13 April 2015 - 02:18 AM

Hello

 

There are a few methods but a cheap method is using bisulfite sequencing.

 

In summary you treat your DNA with bisulfite which changes nonmethylated cyosines to uracils (methylated cytosines remain unaffected). Then you amplify the promoter using PCR and remembering that you use degenerate primers that can amplify methylated and unmethylated DNA. Clone the PCR fragments and then sequence at least 10 clones.

 

There are other more expensive methods that are probably more accurate but as long as you have good controls for your sample bisulfite sequencing is pretty good.   

 

Hope that helps.


Edited by Wunder, 13 April 2015 - 02:24 AM.






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