Inserting Gels into Ettan™ DALT Electrophoresis Units

Two protocols follow, the first for inserting gels into Ettan™ DALTsix and the second for inserting gels into Ettan™ DALTtwelve.

Protocol: Inserting gels into Ettan™ DALTsix

When the electrophoresis buffer has reached the desired temperature, insert the loaded gel cassettes with the Immobiline® DryStrip gels in place.

1. Insert the cassettes into the cassette carrier (Figure 1) and fill any empty slots with blanks.

2. When all six slots are occupied, adjust the buffer level with distilled water so that the level of the diluted anode buffer is at the “LBC start fill” line marked on the unit.

3. Seat the upper buffer chamber over the gels (Figure 2).

Lubricate the gasket and cassette with cathodic buffer (e.g. SDS running buffer) to assist in assembly.

4. Fill the UBC with 1.2 l of 2x buffer. (if not already done in the protocol in section 3.3.1).

5. Using a small funnel, quickly fill the narrow space between the upper and lower buffer chambers with anode buffer or distilled water to the same level as in the upper buffer chamber.

It is important that the anode and cathode buffers are filled to the same height in the Ettan™ DALTsix buffer chambers.

6. Attach and close the lid. Connect the power leads to the power supply.

Inserting the cassettes into the cassette carrier

Figure 1.Inserting the cassettes into the cassette carrier.

Seating the upper buffer chamber

Figure 2.Seating the upper buffer chamber

Protocol: Inserting gels into Ettan™ DALTtwelve

When the electrophoresis buffer has reached the desired temperature, insert the loaded gel cassettes with the Immobiline® DryStrip gels in place.

Gel Cassettes and Blank Cassette Inserts slide much more easily into the unit if they are wet. Distilled or deionized water from a squirt bottle can be used to wet the cassettes and Blank Cassette Inserts as they are being loaded into the unit.

1. Load the unit from back to front (Figure 3).

2. Fit Blank Cassette Inserts into any unoccupied slots.

3. When all 12 slots are occupied, the lower buffer level should be slightly below the level of the gaskets. Pour the diluted (1×) cathode buffer into the upper portion of the tank to the fill line (some of this buffer may drip through the gasket and mix with the anode buffer during the run, but this will not affect performance or results).

4. Close the lid.

Loading the gel cassettes into Ettan™ DALTtwelve electrophoresis unit

Figure 3.Loading the gel cassettes into Ettan™ DALTtwelve electrophoresis unit.

Materials
Loading