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PCR Direct Sequencing

John H McDonald mcdonald at strauss.udel.edu
Sat Jul 16 19:27:40 EST 1994


In article <tay-4.19.000F6501 at bones.biochem.ualberta.ca>,
 <tay-4 at bones.biochem.ualberta.ca> wrote:
          <a bunch of stuff deleted>
>I notice that Pharmacia has a new method that utilizes preferential 
>phosphorylation of one primer and lambda exonuclease. After amplification, the 
>phosphorylated strands are degraded by lambda exonuclease leaving only 
>non-phosphorylated strands for single stranded sequencing. I wonder if that 
>works?

It's not a new technique (see Higuchi and Ochman 1989, Nucleic Acids
Research 17:5865), it works great, and you don't need some silly kit to do
it.  The only sellers of lambda-exonuclease that I'm aware of are GibcoBRL
and Pharmacia, and enzyme from both companies has worked for me.

John H. McDonald
Department of Biology
University of Delaware



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