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GAPDH methylated or not


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#1 march

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Posted 26 March 2009 - 08:55 PM

Hi all,
To see if there is enrichment of methylated gene in IP sample after ChIP,i did PCR.I used primers for a target gene promoter and GAPDH promoter.GAPDH band is of equal intensity in all input and iP samples,and the target gene is mild but can be seen in all.
i)I do not really understand the equal expression of GAP in input sample as well as the IP sample.
ii)I also would like to know if GAPDH promoter is known to be methylated in humans and also how do we check to see if a gene promoter is methylated or not other than the publications.
Thanks

#2 pcrman

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Posted 26 March 2009 - 09:01 PM

By methylation, do you mean DNA methylation? I've never heard of GAPDH promoter methylation in any cells or tissues. What antibody did you use to pull down DNA?

#3 march

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Posted 29 March 2009 - 08:03 PM

By methylation, do you mean DNA methylation? I've never heard of GAPDH promoter methylation in any cells or tissues. What antibody did you use to pull down DNA?

yes,i mean DNA methylation,we used monoclonal Ab against 5-methylcytidine.




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