PAGE gel polymerization

Rob Jordan rjordan at u.washington.edu
Thu Nov 4 15:40:00 EST 1999


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%):

40% acrylamide, 3ml
5X TBE, 2ml
H20, 5ml
10% APS, 70ul
TEMED, 4ul

Now that I think about it does it work better to put the comb in teeth up
for polymerization, then turn it over and use the space between the teeth to
form the actual wells? Hmmm...that's probably the way you're supposed to do
it...

Thanks-
Rob







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