Re- Smeary Touchdown PCR
brett_beitzel at MSMTP.FS.SACI.ORG
Wed Aug 14 08:53:12 EST 1996
Subject:Re: Smeary Touchdown PCR Date: 8/14/96
You can get away with a two step PCR if your annealing temp is 60-70 degrees.
Taq has significant activity at these temps, so it is not necessary to do a
72 degree extension.
How degenerate is your primer? If it is highly degenerate, that could cause
it to give you a smear, especially if your predicted Tm is 60 and you went to
a 55 degree annealing temp during your PCR.
Hope this helps,
brett_beitzel at msmtp.idde.saci.org
Cancer Therapy and Research Center
San Antonio, TX
More information about the Methods