Cracked gels..


Tue May 23 14:42:45 EST 1995


In article <3psvi9$k42 at mserv1.dl.ac.uk>, <grgmj at picr.cr.man.ac.uk> wrote:
> 
> HI - sorry to raise this subject again. The trouble was I didn't pay much attention to the discussion at the time and now a colleague is in trouble with his
> PAGE turning into a mosiac. What are the suggestions for drying high 
> percentage gels? Fixing protocols? Use of glycerol/TCA??? These are the two
> I remember but not in too much detail! I would (and he) appreciate the 
> responses,
> 		Cheers, Michelle
Michelle,
We had similar probs, and glycerol was not the answer. Here were our
changes which allowed for perfect gels:
1. Dont use house vac, very inadequate
2. Do completely immerse your collection flask in ice, it has to be very
cold (you can use dry ice, just make sure you dont freeze the tip closed)
3. Dont use the rigid plastic sheet that comes with some dryers over your
gel
4. Do fix your gel in MeOH/Acetic acid/Water for 30 min before drying
5. Gel should be on Whatman paper, and then covered with Saran Wrap ONLY on
the top (not wrapped all around filter paper)
6. Dry minigel about I hour, big gel 2.  My dryer has a fixed temp, and I
am not sure what it is, but it is hot! (it is a Hoeffer Gelman Jr.)  Also,
some dryers require 2 sheets of Whatman between drying surface and gel
(this is in addition to the whatman on the gel)

We can dry any percentage SDS-PAGE (including 5-20% gradients) with no
problems using this system.



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