PCR from dried sequencing gel?

Bruce Appel appel at uoneuro.uoregon.edu
Tue Nov 15 15:53:48 EST 1994


In article <14NOV94.20401511 at nauvax.ucc.nau.edu>,
schupp at nauvax.ucc.nau.edu wrote:

> Hello,
>         Has anyone tried to perform PCR on bands excised from dried
> Sequencing gels?
> 
> Jim Schupp


The differential display PCR method (Liang and Pardee, 1992, Science
257,967-971) occasionally discussed in this forum requires that
amplification products of interest are isolated from dried, unfixed,
sequencing gels. Soak the gel piece on the paper backing in water a few
minutes, boil it 15 minutes, recover the liquid, phenol/chloroform
extract, EtOH precipitate with glycogen, and put through two rounds of
amplification of 40 cycles each in order to get enough product to clone.
I've had about an 80% success rate.

-- 
Bruce Appel
University of Oregon
appel at uoneuro.uoregon.edu



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