377 Seq 96 lane Upgrade

Jarmo Korkko jarmo.korkko at drexel.edu
Tue Feb 22 09:33:01 EST 2000


We had lots of problems in the beginning, especially with odd-even loading.
The merging was worse in both sides but was happening in the middle also. We
were told that it was salt issue and that didn't make any sense to us (since
we are using columns to purify the sample and we didn't have the problem

At the same time we were trying to start using ABI disposable combs and
personally I think there is something wrong with them....

What works very nicely for us now is the following combination:
Edge or Princeton Separations purification
The Gel Company porous/filter paper combs
All 96 samples loaded at the comb at the same time
Modified water loading protocol (0.1xTBE on the upper chamber, 1XTBE lower
chamber), insert comb, pre-run ~45sec, remove comb, add 10XTBE to upper
chamber to get 1XTBE) and RUN. Works beautifully every time.

Give me a call if you need more detailed information.



Jarmo Korkko, M.D., Ph.D.
Center for Gene Therapy
MCP Hahnemann University
Rm 10118 NCB, Mail Stop 421
245 North 15th Street
Philadelphia, PA 19102
(215) 762 7172
(215) 762 7408 [fax]
jarmo.korkko at drexel.edu


I have recently upgraded my 377 ABI sequencer to 96 lanes. I am having
problems with lanes merging in the first 100bp. There are no salts
involved, and I do a 2 minutes pre-run between loading. Please let me know
If you encountered the same problem, and found the solution.


Miki Rich
rich at tularik.com



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