gel oozing solution posting for autoseq group

Shirley Hall shall at hamon.swmed.edu
Thu Oct 7 09:19:36 EST 1999


Dear David and Group,

I have tired of the gel oozing problem and started reading the board to
find out what people thought.  I don't agree with the idea of that the
process of cleaning the plates is the problem because I've cleaned my
plates the same way since 1990.  I don't agree with the electrostatic
charge because if I ground myself like when I put RAM in the computer the
charge should go away.  What I did realize that this was a problem unique
to my 377 not the 373.  So what was different?  The difference is very
little; molar conc of urea, % acrylamide, thickness of gels and HOW WE PUT
THE COMBS IN PLACE.  For the 373 we rinse the plates off without pulling
out the comb,  pull the comb and wash out the comb area leaving it void of
liquid, put it on the sequencer, scan, put the comb in and then filled with
buffer and pre-run.  For the 377, we rinse the plates off without removing
the comb, pull the comb and rinse out the comb area by filling with WATER
and scraping out all bits of residual acrylamide, FILL THE COMB AREA WITH
WATER TO MAKE THE COMB EASIER TO SLIDE INTO PLACE, then scan, then put the
buffer tank on, then fill with buffer and pre-run.  The whole time the
water is sitting on top of the gel.  The gel, being higher in salt conc,
must be absorbing the water.  I have tried the last couple of days to fill
the comb area only once and absorb it out as quickly as possible with paper
towels (careful not to touch the gel with the towel) and then put the comb
in dry.  Both times no swelling.  I will continue the practise and see if
it stops completely.

Appreciate any feedback if anyone else tries this and the solution works.

Thanks in advance,

Shirley Hall
UT Southwestern Medical Center
shall at hamon.swmed.edu





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