PE ABI 377 Sequencer

Tom Howard (thoward@emory.edu)
Wed, 28 Jul 1999 17:06:19 -0400

Thanks for all of your responses and suggestions regarding my question about
post-sequencing product cleanup in a 96 well plate format. Yesterday I
loaded my first 96 well sequencing gel for the ABI-377 using the Kloehn 8
Channel syringe gel loader recommended by PE. We had some difficulties with
movement of the comb during insertion and removal of the needles at sample
loading. I know that this caused sample leakage into adjacent wells. But
what I didn't expect was that all of the DNA sequences (from every lane) ran
diagonally to the left (if facing gel). The left side of the comb
corresponded to the side that I saw the most leakage. Could a slightly
malpositioned comb cause this deviation of the runs from vertical? If so,
how can I keep the comb tightly in place at loading so that the snuggly
fitting needles don't continue to shift it. Can we tape it at top or maybe
clamp it in some way? Any suggestions will be much appreciated.

Tom E. Howard
Assistant Professor
Department of Pathology
Emory University School of Medicine
1639 Pierce Dr.
Atlanta, GA 30322-3050
Ph. 404-727-3729
Fax. 404-727-3404
E-mail. thoward@emory.edu
Pager: 404-686-5500 (PIC#: 10442)