Help: PAGE sequencing gels

"Alexander Kraev" bckraev at aeolus.ethz.ch bckraev at wawona
Wed Feb 22 12:51:02 EST 1995


In article <wolffie-2002951800380001 at 128.95.13.235>, wolffie at u.washington.edu (John Wolff) writes:
>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

I had the same kind of problem when old rules made me pour gels at least the
day before. Now I usually pour them 30 min before use, prerun for 10 min
without comb, then insert comb and load within the next 30 min. If by
"equilibration" you mean heating up to 50 C, this is not strictly necessary.
If you like to pour gels well in advance ( which also only marginally improve
sharpness of the bands ), you'd better store them with comb removed, the well
drained and wrapped in smth like Saran wrap, without any buffer or wicks.
Hope this helps

*************************************************************************
Alexander Kraev, Ph.D.                 Internet: bckraev at aeolus.ethz.ch
Lab. of Biochemistry III               Phone: 0041-1-632-31-47
Swiss Federal Institute of Technology  FAX:   0041-1-632-12-13
Universitaetstr.16
CH-8092 Zurich
"Some ideas are obscure not because they are complex, but because they 
 are excluded from our circle of comprehension" - Kozma Prutkov



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