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Does anyone have experience with LONG PCR?

Lisa Ruthig ruthig at wista.wistar.upenn.edu
Fri Sep 9 16:06:22 EST 1994


In article <34nl57$fj3 at u.cc.utah.edu>, kelly at thor.med.utah.edu (Kelly
Carpenter) wrote:
> 
> We are attempting to amplify a 16 kb fragment and would like to hear
> about other's experience with using technique, buffer recipes,
> protocols, and kits tried.
> Any or no success using this, we'd like to know.  
> 

    We've been following the protocol in PNAS 91;5695-5699; 1994 (Cheng,
et.al.) exactly, (including using reagents from the same suppliers). 
It's worked better for us than the Barnes protocol; we're getting well 
above 23kb.
    For some primers we have to lower the annealing/extension
temperature, but we haven't had a background problem.
    
Lisa Ruthig
ruthig at wista.wistar.upenn.edu
    



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