PAGE gel polymerization

Peter pxpst2 at vms.cis.pitt.edu
Thu Nov 4 15:58:46 EST 1999


In article <7vsqoc$i70$1 at nntp4.u.washington.edu>, "Rob Jordan"
<rjordan at u.washington.edu> wrote:

> Recently I've started using my own cast TBE page gels instead of precast.
> I'm using an apparatus from Owl to make 8x10cm, 0.8mm thick gels. My problem
> is that the wells aren't formed very well. I'm using a 10 place rectangular
> tooth comb to form wells, and the problem is that when I remove the comb the
> wells aren't nice and straight and rectangular. The sides of the wells
> especially are irregular.
> 
> I'm thinking that the problem is in the polymerization. Here is the recipe
> that I've been using for 10 ml of gel solution (12%):

Did you degass your acrylamide solution (buffer and all?  I filter it
thruogh a .2um filter.  This is especially important if you are using
prebought acrylamide due to the fact that it has a gas stabilizer.
Also, did you allow the gel to sit a good amount of time?  I generally do
not touch the gels for at least 20-30 minutes after casting.

peter pediaditakis




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