Hoefer SE615 - Manual (Page 17)

5
When almost all the acrylamide solution is drained
from the gradient maker, stop the pump and close
the mixing valve. Tilt the gradient maker towards the
outlet side and remove the last few milliliters of mix.
Do not allow any air bubbles to enter the tubing.
6
Add the glycerol displacement solution to the mixing
chamber and start the pump. Make sure no bubbles
are introduced. Pump until the bottom of the caster
is filled with displacement solution to just below the
glass plates, then turn off the pump. Clamp off the
tubing to the red inlet port of the caster.
7
Overlay each separate sandwich with 300 µl of water-
saturated n-butanol or buffer.
Use the same amount of overlay on each gel to
assure that all the gels polymerize to the same height.
Carefully and gently pipette the liquid, taking care not
to disrupt the gel surface.
After polymerization, remove the n-butanol. Replace
it with buffer or add a stacking gel.
8
Allow the gels to polymerize for at least an hour.
To remove the polymerized gels, see “Removing
polymerized gels” on page 17. To add a stacking gel,
see “Pouring stacking gels” on page 15.
•
p14
Note: Isobutanol clouds the
acrylic, making it difficult to see
the gels.