Sequnecing gel problems...

Jernej Jakse j.jakse at Uni-Lj.si
Mon Dec 20 11:26:35 EST 1999


Hello,

We have big, big problems. Using denaturating sequencing gels for
microsatellite and AFLP analysis for almost three years now and till two
weeks ago everything was fine (sigma cote or rain x for non sticky
plate, bind silane for sticky coating, always same urea, acrylamide,
high quality chemicals, using ion resin to remove impurities...). The
gel doesn't polymerize along the spacer. After polymerization (usually
we left gel at room temperature during the night), there is always 1-2
mm of unpolymerized gel solution along the side spacers. And you can
imagine result: buffer from upper chamber is in 5 minutes in the bottom
chamber. Also I  noticed that the gel shrinks a little bit more at the
bottom as in the past. Also tried to change TEMED and APS solution, but
same results. I am applying some grease to help to prevent leaking, but
that is not the solution.
Does anybody have any solution or encountered same problem maybe?

Happy Holidays to all of you, Nejc

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Jernej Jakse
Biotechnical Faculty, Agronomy Department
Center for Plant Biotechnology and Breeding
Jamnikarjeva 101
1000 Ljubljana
SLOVENIA

tel: +386 61 263376
fax: +386 61 1231088

e-mail: j.jakse at uni-lj.si
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