Sequencing gels

Robert A. Anders ander at fermat.mayo.edu
Sat Apr 16 12:29:48 EST 1994


In article <joiner-150494155409 at joiner.med.upenn.edu>,
joiner at pobox.upenn.edu (Michael Joiner) wrote:
> 
> Can anyone tell me how to minimize leakage between adjacent wells when
> using polyacrilamide gels with sharkstooth combs? We seem to have tried
> every combination of prewarming with/without comb, new vs old combs, and
> insertion depth. Whether or not we get leakage still seems to be a random
> occurrence! Does everybody have this trouble?
> 
> Mike

I have this problem occassionally too.  The best thing that I can correlate
it with is a step after the pouring of the gel.  After pouring the gel and
inserting the comb, if I do not put 3 to 4 semi-full 500 ml bottles across
the top edge of plates, I seem to have well leakage. I leave the bottles
there the entire time of the polymerization.  Of course, make sure the gel
if level at this point.  Other suggestions. 1) Don't store plates in a
position that may stress them into deforming. 2)  Make sure plates and
spacers are clean.  3)  Make sure spacers are identical(one isn't for a
wedge gel).
Hope this helps :>
RAA     



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