Amersham Biosciences Hoefer DALT, User Manual, Page: 41 / 70

Hoefer DALT System The DALT Blotting Kit
Amersham Biosciences
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Loading the Cassettes
Work quickly when moving the assembled cassette(s) to the tank to avoid draining the
sponges. Place the tray holding the cassette(s) near the tank, lift out one cassette at a
time, and slide each one into a vertical slot. Do not discard the buffer.
The direction of molecule migration depends on both the characteristics of the sample
and the pH of the transfer buffer. If the species of interest is negatively charged in the
transfer buffer and the stack is assembled so that the membrane faces the anode (+).
Most proteins migrate toward the anode in the Towbin buffer system, independent of
the presence of SDS.
1. Place the support rack for the DALT Blotting Kit into the tank, with the rack
straddling the heat exchanger flutes in the bottom of the tank and the side engraved
“pump” closest to the circulation pump.
2. Lift the cassette sandwich from the tray of buffer and quickly insert it into one of
the sets of vertical slots in the submerged cassette-holding rack.
As many as five gels can be transferred at once in the DALT Blotting Kit. For three
or less gels, use the cassette positions nearest the center.
3. Place the cassettes into the unit so that all gels are oriented the same way, with the
clip side facing up and all anode (–) sides of the cassettes facing the same side of the
transfer unit.
4. After inserting a cassette, tap it a few times to release any air bubbles that may have
been introduced in moving from the buffer tray to the tank. A few bubbles left in
the sponges should not interfere with the transfer.
5. Check the buffer level to make sure buffer covers the cassettes.
WARNING The banana plugs corrode if exposed to buffer during transfer.
Carefully blot away any buffer that falls into the banana plug recesses.
6. When all transfer cassettes are in the tank, place an empty glass gel cassette
horizontally across the tops of the cassettes to anchor them in the tank.
If you are using only one transfer cassette for blotting, place another empty cassette
in a vertical slot to help support the glass cassette anchor.