tips on casting sequencing gel

Stephen R. Lasky Stephen_Lasky at brown.edu
Wed Jul 27 10:13:36 EST 1994


In article <3114a0$8lk at louie.udel.edu>, landel at helios (Carlisle Landel) wrote:

Some text deleted:

> In article <30p5nk$gvp at server.st.usm.edu>,
> Shiao Y. Wang <sywang at whale.st.usm.edu> wrote:
> >pzheng at VAXA.WEEG.UIOWA.EDU wrote:
> >: Put sigmacote on both plates[despite what you have read from sequencing 
> >: protocols]and proceed as usual.

> >One of my students made the same mistake. He couldn't pour the gel because
> >both plate were now hydrophobic and the acrylamide soln wouldn't enter
between
> >the glass plates. So, proceed with caution.

> Let me second Shiao Wang's cautionary note!  If you accidently coat both
> plates, you are in for *big* headaches in getting the gel solution between
> the plates!

Just a comment on siliconizing both plates:  I've been doing that since
about 1982 when I started sequencing using homemade spacers and
sharkstooth combs cut from used x-ray film.  The thinness of the spacers
(about 0.125mm) made it necessary to siliconize both plates, both for
pouring gels without bubbles and for taking the plates apart without
ripping the gel.  I've never had a problem pouring the gels.   I have had
a problem, however, if I didn't wash the plates with comet or something
like that after siliconization:  The gels had a tendency to crawl into the
top buffer during electrophoresis.

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Stephen R. Lasky, Ph.D.       Brown University/Roger Williams Medical Center
e-mail: Stephen_Lasky at brown.edu         LandLine: 401-456-6572
~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~
America may be unique in being a country which has leapt from barbarism to decadence without touching civilization:  John O'Hara
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