We are currently running 64 lanes on each of two ABI 377XL's and are
experiencing a rather odd migration problem with the odd numbered samples.
As most 377 users do, we load the odd lanes first and, after a 5 min.
prerun, finish with the evens and start the normal electrophoresis
conditions of a 2X run. On a whole, when we look at the data the next day,
the odd lanes appear to have less delineated peaks and, once in a while,
have bands that are so "fuzzy" in the fist 100bp, that we are forced to
reload the sample. We have found no correlation with the reagents(i.e.
template and primer) used in these lanes, however, we are consistently
having three or four of these sample results a day. We have attempted
modifying cleanup protocols using several different types of columns, and
have varied the time in which we run the first load in before loading the
even lanes, but, so far, have seen no improvement. I have attached a JPEG
file showing the differences in the electropherograms from samples that are
run on the same gel. Has anyone experienced the same problem or is this
isolated to our lab? I would much appreciate any suggestions on
troubleshooting. Thanks.
-Brian Coullahan
--============_-1288724406==_============
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--============_-1288724406==_============
Content-Type: text/plain; charset="us-ascii"
THE DNA SEQUENCING SERVICE
UNIVERSITY OF ARIZONA
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Skip Vaught- jev@u.arizona.edu
Brian Coullahan- coullaha@u.arizona.edu
Mark Schwartz- schwartm@u.arizona.edu
Amy Raymond- raymonda@u.arizona.edu
--============_-1288724406==_============--