pouring sequencing gels

Bruce Elder elder at CGL.UCSF.EDU
Mon Jun 28 12:28:35 EST 1993



I've been reading all the info on pouring sequencing gels and have to add
my 2 cents. We clamp the plates on both sides but only in the middle, set
the plates on a relatively level elevated platform (pipet tip rack box
works well), tip the plates up at the notched (top) end and "pour" using
a 50 ml syringe. Get a solid 'front' of acrylamide going down the gel and
squeeze spots if you see a bubble form (easier if someone can give you a hand).
It is important to keep a constant flow of solution and not to panic over
the inevitable leaking. The most important aspect seems to be CLEAN plates
so as not to form bubbles. If worse comes to worse, you just start over
having wasted some acrylamide, but saved more time for not having to mess
with all that tape! As you pour, slowely lower the plates back to level
and then clamp at the very bottom and top and insert your comb to make
the well.

Oh yeah, we also have some long spacers that are thinner than our regular
spacers which we use to fish out any bubbles. It is so easy, I am willing
to offer Patricia L. Foster some of our left over rolls of 3M tape if
she still insists on using it!

Bruce Elder
elder at cgl.ucsf.edu



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