If the buffer is in fact wicking over the top as suggested by Sheryl, there is
another option for eliminating the leakage. Introduce a small amount of
Vaseline between the two glass plates after polymerization. I have done this
routinely, and have found that the leakage problem is eliminated. A word of
caution- be sure to clean the plates well! Residual Vaseline on the glass will
cause problems when pouring your gels. As long as you use a very small amount
of Vaseline, and apply it only to the top of the ears, you should have no
problem.
Vergine Furniss
PE Applied Biosystems
Genetic Analysis R&D
_______________________________________________________________________________
Subject: Re: ABI 377 - line in gel file after door opening
From: sherylc@omrf.ouhsc.edu (Sheryl Christofferson) at INTERNET
Date: 1/20/99 11:14 AM
Marcus-
Depending where your upper chamber is leaking, you may be able to stop it. If
buffer is wicking over the top glass tabs, a little blob of polymerized
acrylamide placed on the top tabs of the plate sandwich (not touching the heat
plate behind!) will slow or stop the leak. I pour my gels using a 60 cc syringe
and let the extra polymerize in it. Then I just squish out a little on the top
of those glass tabs. I have been doing this for a year and it works great. Hope
it works for you.
Sheryl Christofferson
OMRF Sequencing Facility
Marcus Macht wrote:
> Dear colleagues,
>
> since some time we have a problem with our Prism 377 DNA sequencer. Since
> our upper buffer reservoir is slightly leaking, we have to control the
> buffer level in the evening. Until December everything was o.k., but when
> we made an upgrade from 48 to 96 lanes, at the beginning we got a line
> which started at baseline level and then turned into a red stripe until the
> end of the scan line. This line occured at the point when we opened the
> door. Since approximately two weeks now, the red stripe turned into a line
> with signal intensity at all for colors for only one scan line as well.
> At first we thought it could be the laser, but it seems to be o.k.
> Has anybody of you already also encountered this problem or knows something
> about its origin? Is there eventually a connection to the "corrupted gel
> file" topic discussed some time ago which I unfortunately did not pursue
> (because I wasn't involved in DNA sequencing at this time ...:-(... )? Any
> help would be appreciated. Thanks in advance.
>
> Yours sincerely,
> Marcus
> ****************************************************************************
> ********
> Marcus Macht
> ZMMK-Servicelabor
> Joseph-Stelzmann-Str. 52
> 50931 Koeln
> Tel.: +49 221 478-6995
> Fax: +49 221 478-6977
> e-mail:Marcus.Macht@uni-koeln.de
> ****************************************************************************
> ********