Help: PAGE sequencing gels

John Wolff wolffie at u.washington.edu
Mon Feb 20 21:00:38 EST 1995


Some of my 6% polyacrylamide gels seem to mysteriously swell while I'm
warming them up:  the gel seems to get squeezed upwards, out of the
plates, and the wells get unusably distorted, and the gel ruined.

I don't think I'm warming them too fast;  we used to equilibrate them at
75 watts, and this has happened at 60 watts.

We recently started using dehydrated TBE buffer instead of pre-mixed
stuff, and this does smell of some kind of osmotic problem.

I've also been experimenting with the polymerization recipe.

I often pour the gels 1-to-4 days before they are used, and this has
happened both with older 1-day-old gels.  I don't think that's the
trouble.  I wrap the comb end with Saran wrap and leave a paper towel
moistened with 1X TBE adjacent to the end to retard drying-out.

Must such gels be stored perfectly flat?  Sometimes I leave them inclined
upwards at a very shallow angle (<5°).

Could it be thermal warping of the glass plates?

I will send electronic chocolate-chip cookies or even Rice Crispy treats
to anybody who can solve this problem for me (they're totally tasteless,
but nonfattening, and it's the thought that counts, right?).



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