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Casting Gradient Gels Hoefer DALT System
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Amersham Biosciences
Figure 11. Priming with
light solution
5. Close both clamps again. All three clamps should now be closed.
6. Add the heavy solution to the heavy (left) chamber (the chamber that is wider at the
bottom) until the liquid level reaches a point about 2 cm below the level of light
solution in the adjacent chamber.
7. Load the side balance chamber with 150 ml dense displacing solution (Figure 12).
The grommet seal and gradient feed tube should prevent leakage of displacing
solution into the caster.
Figure 12. Both chambers
of the gradient maker filled
and the balance chamber
of the casting box loaded
with overlay solution.
8. Open the clamp after the mixer (c), to open the feed tube to the gel caster via the
peristaltic pump.
9. Carefully open the clamp on the light chamber exit tube (a) and turn on the pump
to bring a small amount of solution into the caster.
Light solution
a
b
c
Displacing solution
a
b
c
Light solution
Heavy solution