Hoefer SE215 - Manual (Page 10)

Stacking gel preparation
Add a stacking gel to as many gels as required
either individually to each gel after it is trans-
ferred to the electrophoresis unit (see instruc-
tions with unit), or to the whole stack of gels
while it is still in the caster. To pour stacking
gels in a multiple gel caster:
1
Rinse off the overlay with distilled water. Invert the
casting chamber to drain.
2
Calculate the stacking gel monomer solution volume
(ml): measure the distance, in cm, from the top of
the resolving gel to the notch in the notched plate.
Multiply this distance by the gel width (8 cm), the gel
thickness (cm), and the number of gel sandwiches.
Add the extra volume required for the caster model
used. (Variable Y in Table 2.)
3
Prepare the stacking gel monomer solution, deaerate
it, and add catalyst and initiator. Gauge the amount of
volume required to fill the sandwich to the notch by
adding the stacking gel with a Pasteur pipette to the
first sandwich, which is visible through the faceplate.
Then add the same amount of monomer solution to
each sandwich, working fairly rapidly.
4
Once all stacking gels are poured, insert a comb (at
a slight angle to prevent trapping air) into the front
sandwich, allowing the comb sides to rest on the
spacers. Insert the remaining combs from front to
back in the same manner. Allow a minimum of one
hour for the gels to polymerize. See page 12 for
stack disassembly.
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p8
Important! For optimal gel
performance, cast stacking gels
just prior to use.