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