pouring sequencing gels
Kim Brustoski
kxb38 at po.CWRU.Edu
Thu Nov 30 16:50:21 EST 1995
How about a gel pouring method which does not require the cleanest
plates in the world? The method I use allows some dirt or grease
on the plates because it is so easy to get bubbles out or prevent
them from forming at all. Lay the bottom plate on four tip boxes,
one at each corner, on your lab bench. Place your spacers and anchor them down
by putting a drop of water under them at each end. Pour the acrylamide onto the
plate about 1/3 up from the bottom half. Take your top plate and set it on top
of the bottom plate so that only the bottom edges are touching each other,
making a V-shape. Slowly decrease the angle between the plates by moving the
top plate down. The acrylamide will move up toward the top edge of the two
plates until the two plates are completely together. It is important to create
one gel front because any liquid that is in front of the main front will cause a
bubble to form. If a bubble does form, just raise the top plate until the
bubble is released and then continue lowering to top plate again. It is easy to
prevent bubbles from forming by jiggling the top and bottom plates as you are
lowering or by tapping the top plate as you lower it. Then just put your comb
in and clamp the plates together ( use strong clamps and use 3 or 4 on each
side).
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