Hoefer SE615 - Manual (Page 12)

Pouring standard
homogeneous gels
Table 3. Gel casters and volume required
gel caster required volume gradient maker
(ml) size*
SE615 190–500 SG500
SE675 60 –150 SG100 or
SG500
*Actual size depends on calculated volume.
1
Prepare and degas enough monomer solution to fill all
sandwiches to the level of the notched plate.
The following formula allows extra solution to fill the
space between the sandwich and the chamber:
No stacking gel. Fill solution to just below the top of
the notched plate. If air pockets form, remove with a
pipette or syringe. Introduce a comb, at a slight angle,
into each sandwich, taking care not to trap air under
the teeth.
Stacking gel. Fill solution to 4 cm below the top of the
rectangular glass plate. This height allows 1 cm of
stacking gel below the wells. Pour the gel and apply
an overlay. After the gel is set, prepare the stacking
gel as described on page 15.
2-D electrophoresis. Fill solution to about 1.0 cm
below the top of the rectangular glass plate. This
height allows 4 to 5 mm of space for the IPG strip
and an agarose seal. Overlay the separating gel as
described in step 3.
Warning! Acrylamide is a
neurotoxin. Wear gloves
when handling acrylamide or
polyacrylamide. Wear a dust
mask when weighing acrylamide
or preparing acrylamide stocks.
•
p9
Note: The 1.1 multiplier in this
formula assures enough solution
to fill dead volume. Monomer vol. = height × width × spacer × total number of × 1.1
(ml) (cm) (cm) thickness (cm) gel sandwiches