PAGE gel polymerization

George Rutherford gruther at bilbo.bio.purdue.edu
Mon Nov 8 16:03:14 EST 1999


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


> 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


Some snippage....

Unless your lab is really cold, those gels must be polymerizing like a
shot. Although I make up a larger volume at a time, I use the equivalent
of 30ul of 10% APS and 3ul of TEMED and my gels polymerize in 15-20
minutes, although I don't pull the comb for at least an hour. I rarely
have problems with bent wells, but if I do, a small gauge needle can be
used to straighten them out. BTW, how much Bis are you using? I use 0.8%
in a 30% stock solution. High levels of Bis can affect the extent of
crosslinking and the rate of polymerization.
Good luck,

George




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