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Low MSP sensitivity - (Oct/15/2008 )


I have a problem of low MSP sensitivity.

I am getting only primer dimer and not the specific band in methylated gene specific PCR, which I should get according to the referece.

I am getting clear band with universally methylated genomic DNA I had arranged.
In the reference, nested PCR was used to overcome the small amount of methylated gene existing in the genomic DNA.

So I think I should raise the sensitivity of MSP.
What should I try first and next in this case?

Adding DMSO? higher MgCl2? more template?

-awhite0-

adjusting/optimising your PCR conditions to suit your own conditions would be the first thing to do before delving into DMSO and Mg concentrations.

Nick


-methylnick-