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problems with Bisulfite PCR


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#1 vaidhi

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Posted 12 February 2012 - 09:40 PM

Hello,

I'm new to the the world of methylation and am trying to amplify CpG islands after bisulfite conversion. But when i run the PCR products on a gel i get smears and they aren't even in the correct range. I designed my primers using meth primer and when the first PCR didn't work i tried lowering the extension temperature to 63 C ( first i used 72 C).

Any suggestions would be greatly appreciated.

Edited by vaidhi, 12 February 2012 - 09:41 PM.


#2 methylnick

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Posted 29 March 2012 - 02:03 PM

most liekly a primer design problem, I would try to redesign them again, maybe using MethylPrimer Express which is freely available.

All comments and communication are my own personal ones, and are not tied to any of my affiliations. 





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