Re- Smeary Touchdown PCR

Brett Beitzel 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
Cancer Therapy and Research Center
San Antonio, TX

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