Hoefer SE400 - Manual (Page 19)

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2.2 Acrylamide gel preparation
Table 1. Approximate monomer solution volume
required for a single gel
Gel thickness (mm)
Model 0.75 1.00 1.5
SE400 15 ml 23 ml 30 ml
SE410 23 ml 34 ml 45 ml
2.2.1 Resolving gel
1
Prepare the monomer solution and pour the gel.
Prepare the required amount of monomer solution,
deaerate, and add the initiator and catalyst just prior
to pouring the gel.
2
Pipet the solution into one corner of the sandwich,
taking care not to introduce any air bubbles. See
below for the appropriate solution level:
No stacking gel (Continuous system). Fill solution to
just below the top of the upper plate edge. If bubbles
are trapped, remove with a pipet or syringe. Introduce
a comb (at a slight angle) into each sandwich, taking
care not to trap air bubbles under the teeth.
2-gel sandwich. Pipet the solution into both sand-
wiches, filling each to the same level below the
notched edge.
Stacking gel. Fill solution to 3–4 cm below the top of
the glass plate. This height allows 1 cm of stacking
gel below the wells. Pour the gel and apply an overlay
(see step 3). After the gel is set, prepare the stacking
gel as described in the next section.
2-D electrophoresis (Discontinuous system). For the
second dimension resolving gel, fill solution to ~1.0
cm below the top of the glass plate (leave extra space
for a stacking gel, if required). One centimeter allows
enough space for the first dimension IPG strip or tube
gel and an agarose seal. (While transferring, take care
to avoid trapping air between the tube gel and slab
gel; seal the tube gel into place with agarose in elec-
trophoresis buffer.)
Note: Appendix A on page
29, lists recipes for the
Laemmli gel system.