C.B.S. Scientific DH Series - Manual (Page 15)

C.B.S. œ Scientific
15
Dedicated Height Sequencer
6. Before the acrylamide polymerizes, insert a comb into the top of the gel. The available types
of combs are: rectangular tooth combs, a casting comb the same thickness as a sharktooth
comb or the back side of a sharktooth comb as a casting comb. If you are using a rectangular
tooth comb, insert to edge of backing. If you are using a separate, one well casting comb with
a sharktooth comb, insert casting comb to edge of backing. If you are using a sharktooth
comb, insert the back side of the comb to the black lines (depth indicators) printed on the
comb (see figure below). The depth indicators are pre-determined and specific to each well
configuration.
7. If you decide to clamp “over” the casting comb, to ensure a preciously defined gel thickness,
use clamp cat. # GPC-0004. It is important the spring clamps be placed only over the
comb (and side spacers), clamping unsupported glass will distort the thickness of the
gel. See illustration above.
8. Cover the top and bottom of the gel plate assembly with plastic wrap so that as little air as
possible can contact the gel. Allow the gel to polymerize for 30 minutes to 1 hour depending
on the width of the gel poured. The gel can be poured the day before the run and stored at
room temperature for a maximum of 48 hours, if wrapped with a wet paper towels and then
with plastic wrap.
9. Leave the glass plates in their near-horizontal position until the acrylamide has polymerized.
After polymerization remove the clamps. Rinse surface of the glass plate assembly with de-
ionized water to remove any excess gel. Carefully pull the comb from between the glass
plates. If using a rectangular comb skip to step 4.1-2. Rinse the top of the gel with
electrophoresis buffer to remove any unpolymerized acrylamide. Rinse the comb with
de-ionized water.