pouring sequencing gels

John McElver mcelverja at phibred.com
Mon Nov 27 20:22:00 EST 1995


Re: Re:pouring sequencing gels

  I vote for slide casting while flat, I've used a modified Pharmacia
MacroMould to cast gels for an ABI 373a sequencer, as well as the
Pharmacia system it was originally made for.  It it one heck of a lot
easier and faster than taping.  I've done it this way since '89 or so,
until I moved here and we bought a Pharmacia ALF.  It has it's own set-up.

Basically it works like this (hopefully this'll come across OK):

Tape spacers in place with small pieces of 2 sided tape.

Get the plates on the apparatus as below.
                 __________________
___________________                }glass plates

Pour the gel solution at the intersection of the plates.  Allow the gel
to flow to the "bottom" by capillary action.

Begin to slide the top plate to the left:
                  <---
        __________________
___________________ 
while pouring more gel at the "leading edge."

Continue to pour as you slide the top plate to the left.  The apparatus
supports the top plate to keep it level and allows it to slide to
the right readily.  As long as a good "bead" of gel solution is kept at
the leading edge, it is impossible for bubbles to form, unless the plates
are filthy.

When the plates are registered, clamp and insert the comb.  Oh, yeah, the
pouring surface pretty much has to be level!

I think Owl Scientific might sell a variation on this theme, and a
homebrew variation was discussed in a Biotechniques a couple of months
ago.

Basically, I don't know anybody who has gone back to tape after casting
this way.

John



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