Electrophoresis Using Buffer Chambers - Pharmacia Biotech Multiphor II User Manual

5. Operation
5.3 Electro-
phoresis
using buffer
chambers
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staining and preserving methods recommended for ExcelGel SDS (See
Section 4.1) work well with CleanGel.
This section describes the running procedure when using buffer chambers.
To place the electrodes in the buffer chamber, remove the cooling plate
from the buffer tank.
To reduce the effect of electrolysis products during the electrophoresis, the
electrodes should be positioned as far as possible from the wicks.
Therefore, when performing electrophoresis across the large cooling plate,
the electrodes should be placed in the grooves in the wall closest to the
centre of the unit. The wicks lie at the outer edge of the buffer chamber.
Place the cathode electrophoresis electrode in the left buffer chamber and
the anode in the right buffer chamber.
Fill each chamber to the moulded line (indicating 1 liter volume) with
buffer solution. (When running 120x250 mm gels, pour 1.2 liters of buffer
into each chamber to ensure adequate buffer contact with the wicks.)
Replace the cooling plate, making sure that the electrode socket connectors
lie to the front and that the connecting cable is clear of the feet on the plate.
Disconnect the anode bridging connector for isoelectric focusing and
connect the electrodes to their respective pins.
When performing immunoelectrophoresis or agarose electrophoresis, center
a small (84x94 mm) glass plate on the dry cooling plate and attach a strip
of tape along the width of the cooling plate in alignment with the edges of
the glass plate. These stop the small gels from shifting during application of
the wicks and ensure that they are centred between the two buffer
chambers during electrophoresis.
Switch on MultiTemp III thermostatic circulator and set the desired
temperature (normally 10 °C for PAGE or agarose electrophoresis and 15
°C for SDS-PAGE) 15 minutes before starting the experiment.
To ensure efficient heat transfer from the gel during electrophoresis, a
uniform layer of a non-charged insulating fluid is applied under the gel. To