Hoefer SQ33 Sequencer - Manual (Page 15)

3. SQ33 Operating
Instructions
3.1 Regent Preparation and Gel Volumes
For reproducibility and uniform polyacrylamide
cross-linking we recommend deionizing, degas-
sing and filtration of acrylamide gel solutions
prior to use. Acrylamide solutions should be
stored in a cool, dark environment such as a
refrigerator and allowed to reach room tempera-
ture prior to pouring. Avoid exposure to heat
and sunlight.
Polymerization conditions should be adjusted
to effect polymerization within about 5 – 10
minutes. Test a small volume in a vial prior to
pouring the gel. As a rough guide 100 ml of
degassed 6% acrylamide gel will set in about 5
minutes at room temperature when gently mixed
with 450 μl of freshly prepared 10% (w/v)
Ammonium persulfate plus 200 μl TEMED.
The setting time increases to about 10 minutes
if the TEMED volume is reduced to 100 μl and
to approximately 15 minutes with 75 μl. The
amount of catalysts may need to be reduced
under warm conditions. Do not pour under
direct sunlight.
The SQ33 Sequencer, using 0.35 mm thick
spacers, requires approximately 60 mls of
acrylamide solution.
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