PCR-PROPDUCT PURIFICATION

J.J. Baker mbjjb at s-crim1.dl.ac.uk
Tue Nov 15 07:40:20 EST 1994


sl6dp at cc.usu.edu wrote:
: Going through the literature I have found that it is possible to ream-
: plify PCR products directly from the band after selection on a low-melting
: agarose gel. I am not sure if I understand it: it is really possible
: to run the PCR with a portion af the agarose band, without even
: taking the DNA out????. Anyaway, which is a good method for PCR product
: purification if I want to run a secon PCR with another primer?, I mean
: no comercial kits...Thanks

Yes it is! Though I can't remember the exact referencefor this technique,
this is the method I use; I pierce the bands of the gel with a wide bore
needle and then put the needle into the reaction buffer for PCR then leave for 
about ten minutes or the length of your tea break at room temperature
Then simply take out the needles, add your Taq and run for 5 more cycles
than usual.

So far this method has only failed me once and that was with a very faint
band so obviously the amount you load on the gel in the first place is
quite important.

Trust me, it works!

Jon Baker

Centre for Molecular Biology
Dept Chemistry
The School of Pharmacy
29/39 Brunswick Sq
London
WC1N 1AX   UK



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