Pouring sequencing gels w/o leaking

Klaus.Matthaei at anu.edu.au Klaus.Matthaei at anu.edu.au
Tue Jul 6 21:23:12 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

Yep I agree with all of the above.  The Biorad IPC's are the way to go and
I have been using them for about 5 years.  I do not use a plug but rather
push a piece of 3mm into the bottom of the plates, clamp with an electrical
clamp and pour at a 30* angle without an acrylamide plug.
Bubbles you only get when the plates are not clean.

Cheers, Klaus
Klaus Matthaei
Gene Targeting
The John Curtin School of Medical Research
The Australian National University
E-mail: Klaus.Matthaei at anu.edu.au

"I know that you think that you think that you understand what I am saying."
"But what I am saying is not actually what I mean."

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