Odd problem with sequencing gels
The doctor is IN.
aarment at desire.wright.edu
Sun Jan 23 15:17:59 EST 1994
I am having an odd problem with my sequencing gels that no one around
the university can offer an explanation for; thus, I appeal to the wisdom of
the bionet. My problem is this... when I run a sequencing gel, everything
begins just fine. I like to run around 1700V and 40W. The gel preheats, I
load it, and then trouble begins. Over the course of the run, the wattage
begins to decline (and with it the ampage), although the voltage remains
constant. After about two hours, the wattage is down to 25W, and the gel has
cooled to the point where it is useless for sequencing.
I have changed gel buffer systems (TBE and TEB), changed the way I pout
my gels, changed the percantages of acrylamide, recirculated the buffer,
and completely changed the buffer; all to no avail. The power
supply appears to be working correctly. It runs other gel boxes without this
porblem. I can generate current across the upper well but NOT through the gel.
Does anyone have any ideas and/or suggestions as to what is going on?
Wright State University
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