Acrylamide Gel setting too quickly -- IDEAS??
mdpjr at cc.newcastle.edu.au
Tue Jun 4 10:59:34 EST 1996
macnevin at ix.netcom.com (Brian MacNevin) wrote:
>Well, when I mix up my acrylamide gel solution and begin pipetting it
>between the glass plates for electrophoresis, I have found that it
>starts to set before I can get it evenly dispersed in the plates.
> My first thought is that this might be due to the temp of the
>solution. It seems to me that I don't have this problem if the
>solution is allowed to cool a little before adding the final solution.
>I was wonderin if anyone else has had these problems and if it really
>is due to the temp of the solution?
Seems like too much APS and/or TEMED. Start by reducing the TEMED.
Set up little test lots in eppendorf tubes and see how long they take
to set. Something like 10-20 min should be fine.
However, we approach the problem by cooling the acrylamide solutions
on ice. We pour gradients and to prevent any chance of it setting in
our gradient mixer or tubing, we and many others, find that keeping
the gel solution on ice prior to pouring significantly slows down
polymerisation rates. And old and well-known trick... Should not be
necessary for single-percent acrylamide, although we do it there too -
out of habit more than anything.
Phil Newcastle Australia
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