Pharmacia Biotech Multiphor II User Manual page 75

9. Multiphor II application kits and accessories
Prepare the agarose solution in a water bath and cool to 75 °C before
adding the carrier ampholytes. Take the mould out of the heating cabinet.
Slowly fill a 20 ml syringe (with flexible tubing attached) with the molten
agarose solution. Avoid drawing air bubbles into the syringe.
Hold the mould at a 20° angle. Inject the solution between the glass plate
with spacer and the film. The gel solution will flow into and fill the mould
by capillary action. As the mould fills, lower the angle to decrease the flow
of the gel solution. When completely filled, place the mould in a horizontal
position and allow the agarose solution to gel for 15 minutes at room
temperature.
Before opening, remove the excess gel from the ends of the mould with a
scalpel. Remove the two FlexiClamps. Stand the mould on one long side.
Grasp the GelBond film and glass plate with spacer with one hand and the 3
mm glass plate with the other. Pull apart gently. Remove the 3 mm glass
plate and then the glass plate with spacer from the gel. Place the gel in the
Humidity Chamber at 4 °C for at least 1 hour. The gel is now ready for use.
Alternatively it can be stored in the Humidity Chamber at 4 °C up to three
days.
Before use, lay a sheet of filter paper on the gel surface. Check that there are
no air bubbles trapped. Leave for l minute and then carefully peel off the
filter paper. The gel is now ready for IEF.
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