Hi,
Several things.
You do, of course, need to make sure the combs and spacers are clean
and free of dried acrylamide.
Probably no two combs are identical in thickness. We had the best
results polymerizing the gels against the backside of the same
sharkstooth comb used to form the wells.
If an old comb has been bent or creased, it is nearly impossible to
make it perfectly flat again, therefore the teeth will be loose when
the comb is flipped over to form the wells. For the same reason -
don't use bent spacers.
Less likely but the comb might be thicker at one end than the other -
thus be sure you simply turn the comb over and don't flip it end for
end.
Make sure the clamping frame is in good condition and that the clamps
are working as firmly as always.
If the gel crosslinking is lower, it will be more flexible and not
hold the plates together as tightly. Make sure the acrylamide is not
too old and that polymerization time has not increased.
Look forward to hearing what the fix is.
Best regards,
David Bishop, Ph.D.
Director, DNAcore
Mount Sinai School of Medicine, NYC
>Hi there,
>>We're experiencing a problem with the sharkstooth combs for the ABI
>377, and any suggestions would be much appreciated.
>>We're finding that our wells have started leaking when we load the
>gel, and the problem seems to be that the well formed is perhaps too
>wide, making the teeth loose in the well. We don't get acrylamide
>in the well, so it doesn't seem to be a problem with the clamping.
>>New combs work fine, which suggests that there might be some build-up
>on the older combs?
>>Has anyone experienced this? If so, any suggestions on the best way
>to "strip" the combs?
>>Many thanks,
>-Samantha Donninger
>>>------------------------------------------
>MRC Human Genetics Research Unit
>Division of Human Genetics
>Department of Clinical Laboratory Sciences
>University of Cape Town
>>email: st at cormack.uct.ac.za>ph: (+27 21) 406 6456
>fax: (+27 21) 488 0906
>>---
--
David F. Bishop, Ph.D. | EMail: david.bishop at mssm.edu
Professor of Human Genetics | Phone: (212) 659-6795
Dept. of Human Genetics, Box 1498 | FAX: (212) 849-2508
Mount Sinai School of Medicine |
1425 Madison Avenue, NY, NY 10029 | www.mssm.edu/genetics/home-page.html
---
