Selective PCR amplification

Ted Michelini tedm at darkwing.uoregon.edu
Tue Oct 22 12:19:09 EST 1996


In article <32315370FB5 at oreo.uottawa.ca>, djohnson at OREO.CHEM.UOTTAWA.CA
("Doug Johnson") wrote:

Doug,
   An air cycler (Idaho Technology) set to extend for a sparse few seconds
may give you more short product, (I always seem to get much more small
product than long!) I guess it all depends on what you want. The extension
of Taq is supposedly 100-300 bases per second, or something like that, so
you have to be quick. Good luck.

regards,
Ted Michelini
Institute of Molecular biology
University of Oregon


> We are trying to clone several closely related sequences via PCR 
> using the same primers. The products are different sizes, from 300 to 
> 600 bp. When we amplify and clone, we have to screen many recombinant 
> before we can find the smaller ones. Is it possible to control the PCR 
> conditions e.g., by lowering the time of extension, in order to selectively 
> amplify the smaller products?
> 
> Any suggestions greatfully considered.
> 
> Doug Johnson
> Dr. D.A. Johnson
> Plant Molecular Biology
> Department of Biology
> University of Ottawa
> 30 Marie Curie
> Ottawa, Ontario
> K1N 6N5
> 1-613-562-5800 ext 6347
> 1-613-562-5486(FAX)



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