Using DS DNA as a primer
groi9663 at uidaho.edu
Sun Jun 28 14:49:16 EST 1998
Well here is one we have been struggeling with...
A fellow Grad Student of mine is attempting a PCR reaction in which one
of the primers is actually a DS DNA fragment which had been purified
earlier. Upon running the rxn we get various non-specific products, and
a REALLY low amount of the expected size product, not enough to be of
The Pol we are using is vent, we have tried w/both MgCl2 and MgSO4,
various conc. of course.
And the DS primer is ~325bp in length. We have tried to vary the
Tanneal within a 15 degree range.
So my question is, has any one tried using a DS DNA frag as a primer,
and if so, how did ya get it to work.
Any help or insight would be greatly appreciated. Thanks in advance
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