sequencing plates problem

HARDIES at THORIN.UTHSCSA.EDU HARDIES at THORIN.UTHSCSA.EDU
Fri Jan 13 15:34:13 EST 1995


To: methods-and-reagents at net.bio.net
From: parmentf at merck.bch.umontreal.ca (Fabrice PARMENTIER LESAGE)
Subject: Sequencing plates problem
Date: 13 Jan 1995 11:05:30 -0800
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Fabrice Parmentier Lesage writes:

> I am using S2 Sequencing System BRL 0.4 mm. I have to dry the gel on the 
> plate. When I disassemble the gel sandwich, it doesn't stick on the 
> proper plate (even if the other plate has been siliconized). Even if the 
> gel sticks on one plate, it does not stick enough. When I wash the gel 
> it often slips away from the plate and the same thing happens when I try 
> to dry it with a hair dryer.
> If you have any ideas how I could treat the plates before pouring the gel 
> please let me know.

Sounds like both of the plates are siliconized.  If you scrub the
plate you want the gel to stick to thoroughly with a NONabrasive
cleanser, it may solve the problem.  This won't take the silicon off,
but it will make it stick a little better.  The plate that's supposed
to be siliconized needs to be resiliconized every 2nd or 3rd use or
else it wears off (at least for the sigmacote I use).  I assume BRL
hasn't done anything funny to the glass, but it wouldn't hurt to
call and ask their advice.

If by "washing" the gel you mean it flows off the plate when you pick
it up from a water or fixative bath, you can save that by putting 2
strips of Whatman paper about 3 inches wide by the length of the gel
into the bath on top of the gel over where the spacers were.  Catch
the paper under the thumbs of your gloves when you pick it up and the
gel will come out without sliding off even if it is completely
dislodged.  Lots of people don't soak or fix the gel at all now; see
the FAQ for details.

I don't understand the part about the hair dryer.  If you're not going
to transfer to paper and dry, then just put plastic wrap on it and 
expose direct.  From prior comments in this group, apparently people
even do that for S35, although it must cut the signal down some.

Hope this helps.
Steve Hardies, Dept. of Biochemistry, Univ. of Texas HSC at San
Antonio
Hardies at uthscsa.edu






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