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Hoefer DALT System Preparing the DALT Tank for Electrophoresis
Amersham Biosciences
25
gels, up to the tank maximum. The empty slots seal themselves and require no blank
cassette. Since the filled tank is too heavy to empty directly, use a siphon pump to
remove buffer from the tank. Between runs, rinse the tank with water.
1. Two or three hours before the beginning of a run, fill the tank with the appropriate
volume of de-ionized water. Use tape to mark the level of the liquid on the outside
of the tank for future runs.
Always use water near room temperature. Never fill the tank with liquid at a
temperature very different from the tank itself.
I
MPORTANT
Fill the tank with water before turning the pump on. The pump is not
self-priming and can be damaged if run dry.
2. Turn on the pump after the tank is filled.
At first, the pump blows some bubbles through the circulating flutes in the tank and
then establishes a vigorous circulating action in the tank. If the pump doesn't
“catch” and no circulation is observed, you have an airlock in the pump. Quickly
turn off the pump. Wait a moment for the air to bubble out through the small
“bypass” tube that comes up from the pump outlet and enters the back of the tank
about halfway up, then restart the pump. You may have to do this a few times.
Once the pump is circulating, leave it on.
3. Weigh out the correct SDS electrophoresis buffer mixture. (See page 42.) Add the
dry powder directly to the water in the center chamber, distributing it evenly from
front to back.
The pump circulates the contents enough to dissolve the solids in about 1-2 hours.
Some solids may lie on the bottom for awhile, but they gradually disappear. You
may want to make a number of these buffer packets in 1-liter plastic bottles and
have them on hand.
4. Once the buffer is dissolved, use a Phillip’s head screwdriver to loosen the white,
plastic retaining screws on the exterior of the DALT Tank that hold the barrier
combs in place.
Raise or remove the two barrier combs to mix the total DALT Tank contents with
the dissolved contents of the center chamber. (See Figure 14 on page 26.) You may
tighten the retaining screws to secure the barrier combs in the raised position while
the buffer circulates throughout the tank.