ELECTROPHERISIS

Laura Moen lmoen at odu.edu
Wed Sep 16 15:33:35 EST 1998


I agree with the earlier post - use extra clamps up at the top of your
sequencing set up to hold the plates tighter.  Put these clamps on _after_ you
put the comb in.  The suggestion about the thickness of the comb is good too.  I
don't grease my comb, but I don't know if that has any effect on your sample
leakage or not.  One more thing - once and awhile you get a comb that has a few
wells where samples just leak from time to time.  I don't know why that is, but
we mark the teeth around those wells to avoid loading them.  Also, check your
spacers for your gel - they may not be the thickness you think they are.  Good
luck.  Laura Moen



jose manaligod wrote:

> This is once the gel is placed into the electropheris box and clamped into
> place.  We then place the comb into the gel and after testing the amount a
> well can hold with a -c, we then load accordingly.  It then leaks from
> behind the comb into the well beside it.  In article
> <jmmana-1609980830560001 at 128.163.85.156>, jmmana at pop.uky.edu (jose
> manaligod) wrote:
>
> > We are running 6% Acrylamide gels using .4mm spacers and combs.  We are
> > having a lot of problems with our samples leaking out of the wells into
> > the one beside it.  After our PCR we use 8ul of the PCR sample and 3ul of
> > dye.  We greese the comb with vaccum grease.  Has anyone had this problem,
> > if so do you know how to prevent leaking?
> >
> >
> > Magie Chokloikaew
> > University Of Kentucky
> > Dept. Otolaryngology/Molecular Genetic Research
> > 800 Rose Street
> > Lexington, KY 40536-0084
> > 606-323-8287office
> > 606-2575096 fax
> > mchokl at pop.uky.edu






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