Pouring sequencing gels w/o leaking

dnicker at vax.oxford.ac.uk dnicker at vax.oxford.ac.uk
Tue Jul 6 18:05:26 EST 1993


	And yet another dives into the flurry of activity on this subject. . . 

Okay, here's my 2 cents. . . I am surprised that there has been (so far as I
know, and I may have missed the beginning of this thread) no mention of the
latest apparatus being marketed by Bio-Rad.  As a total amateur I began using
their setup and had gel-pouring mastered very quickly.  Their method involves
clamping the plates with spacers, and casting a plug bafore pouring the main
gel.  The clamped setup is placed vertically (free standing for the excitement
inclined) into a "gel casting tray" into which 30 ml of gel sol'n is poured. 
With enough TEMED and APS, this sets within 5 minutes.  No muss, no fuss, and
you have a strong plug.

Only stumbling block I encountered is that the main gel MUST be poured on a 30
degree incline from horizontal, not when the setup is vertical as Bio-Rad
suggests.  This sometimes will generate enough force to burst the plug. 
Since making this modification, have never had a leak :)

As for the bubble dilemma. . . with clean plates and slow steady pouring this
has never been a problem either.  Maximum of 4 bubbles in over 30 gels poured,
and I just work around them.  Now though, am looking into the wooden phage
models to burst them as they form, and may even try the assorted hooks and
other such "bubble catchers" mentioned here :)

I must mention that I am in no way affiliated with Bio-Rad, _________ <-- fill
in the rest of the standard disclaimer here please =)

Darren Nickerson
Inorganic Chemistry Laboratory
University of Oxford
England



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