using the meth primer - (Feb/16/2009 )
I am trying to get the primers for the promoter region of usf1. I have picked the sequence in NCBi and pasted in meth primer... I got five options, but the problem is that the amplicons for M and U primers have the same numbers of pb e.g : 150 e 152 pb. How will I distinguish them in poliacrilamide gel???
Iam new in this subjet and iqam trying to find the way.
It's OK if you would not mix up your M and U DNA templates. For example, do the M template PCR on day 1 and the U template on day 2. Or if you use methylight Real-time PCR with two taqman dyes. But I would try to pick another primer pair, just in case you might mislabel the tubes.
If you do fecal DNA hypermethylation study, please PM me and I can give you some idea of getting rid of the PCR inhibitors from your DNA preps.
you would run the M and U primers in separte PCR reactions and then run them in different lanes on a gel, doesn't need to be PAGE.