Using DS DNA as a primer

Linda Western western at cepheid.com
Mon Jun 29 14:13:23 EST 1998


Why are you using a ds primer, and why is it so long?  I've been doing PCR
for over 10 years and have never heard of this.

Kajetan Groicher <groi9663 at uidaho.edu> wrote in article
<35969E3C.48F8 at uidaho.edu>...
> Hello All-
> 
> 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
> use.
> 
> 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
> 
> Kajetan
> 



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