Preparative acryl amide gel electrophoresis

Peter pxpst2 at vms.cis.pitt.edu
Fri Jun 11 18:20:42 EST 1999


In article <OLTpudFt#GA.202 at ntawwabp.compuserve.com>, "Gys de Jongh"
<GysdeJongh at csi.com> wrote:

> Could it be that the dna reacts chemically with the acryl amide monomer ?
> or with the oxygenradicals from the persulfate/temed ?
> or with the glycylamide formed by the above two ?
> Has anybody used acryl amide gels for preparative purposes ?

If you speculate that the arcylamide or reagents used in the
polymerization are involved then you can do what protein chemist have done
for a while when PAGE is used in protein sequencing:

1) Allow gel to sit overnight before running
   > Free radicals are not very stable and their concentration will
disipate exponential over time.

2) Prerun your gel for about 20% of the total expected gel run time.

   > Any remaining ions can be pushed ahead of your dye front, insuring
that they are not around when the DNA comes through

regards,
Peter



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