Help!PCR problems.

Michael Cooley szcooley at dale.ucdavis.edu
Fri Feb 17 17:19:10 EST 1995


sl1nn at cc.usu.edu wrote:
: I am having problems with PCR amplification of a small fragment of cloned DNA.  I
: I have designed my own primers, using Oligo 4.1, and according to the program I
: am using the right temperatures for the primers. I have varied tha conditions
: several times with the same results.  The negative control (without
: template) of my two primers always shows a 250 bp band.  Both primers are
: 22-mers and the dimers would not be that large. In additon, when I have the
: template present I am only seeing the same band. I have checked the primers for
: contamination using HPLC, and they check out at the right size and with only
: one peak.  Does anybody have any idea why I am getting this result?  I'm going
: crazy trying to fix the problem.   Thanks in advance.

Is it possible that the fragment is coming from DNA known to contaminate 
most TAQ enzyme preps?


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