Pharmacia Biotech Multiphor II User Manual page 91

9. Multiphor II application kits and accessories
If the channel is blocked with polymerized gel solution, disassemble and
clean the Gradient Maker as described below.
Close the channel by pushing the valve to closed (c) position. Check that the
pinch cock is closed.
Prepare the dense and light solutions.
Note: Gloves must be worn to protect the user from contact with toxic
acrylamide solution.
Fill the reservoir with the light solution. To fill up the channel with light
solution, open the valve carefully until a drop emerges in the mixing
chamber and close the valve immediately. If light solution has entered the
mixing chamber, use a pipette to transfer it back to the reservoir.
Fill the mixing chamber with the dense solution. Center the mixing chamber
over the magnetic stirrer. Check that the gradient maker is levelled. Place the
prepared mould on a levelled table and insert the end of the tubing between
the glass plates in the centred groove. The tubing should lie against the gel
support (glass plate or film), not the glass plate with U-frame. The slot
former will otherwise disturb the flow of the solution.
When preparing a linear gradient, place the compensating stick in the
reservoir. For good linear gradient formation, the difference in height
between the Gradient Maker outlet and the end of the tubing inserted in the
mold should be about 5 cm for a total gel volume of 15 ml and 7 cm for a
total gel volume of 60 ml.
When preparing an exponential gradient, insert the plunger in the mixing
chamber until it is 0.5-1 cm above the level of the solution.
Turn on the magnetic stirrer and check that the bar rotates smoothly,
creating a small vortex. Set the stirrer to 250 rpm for 15 ml gels and to 300
rpm for 60 ml gels.
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