Pouring sequencing gels w/o leaking

[Klaus.Matthaei at anu.edu.au]

>        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


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
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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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