Sequencing gel combs (leakage)

Warren Gallin wgallin at gpu.srv.ualberta.ca
Tue Mar 29 14:14:03 EST 1994


In Article <CnFMtE.KM8 at usenet.ucs.indiana.edu>,
jgraham at bronze.ucs.indiana.edu (the End) wrote:
>
>Great responses. 
>
>One thing that seems to be coming up is the desirability of "clamping"
>over the comb during polymerization (as well as running). This would 
>seem counterproductive in that it may hold the plates closer together
>while the gel sets than the plates will be during the run (if unclamped).
>
>This would seem to increase leakage, however, not necessarily.
>
>Anyone ?
>
   I have found that clamping the plates over the comb during polymerization
is essential.  If you don't, you can get a thin layer of acrylamide between
the comb spacer and the plate.  Then when you try to insert the sharks tooth
comb, all your wells get fouled with the extra polyacrylamide.
   As for clamping while loading and running, I hadn't thought of it but it
sure sounds reasonable, and it doesn't cost anything to try it.  Most of the
problems that I have had with spillover, though seem to be due to the sample
going under the comb tooth, not around it.  Maybe clamping will force the
gel up a little more tightly to the bottom of the tooth.

Warren Gallin,
Department of Zoology, University of Alberta
wgallin at gpu.srv.ualberta.ca



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