Hoefer SE615 - Manual (Page 16)

Pouring gradient separation gels
1
Prepare and degas the two monomer solutions for the
gradient maker.
Add glycerol or sucrose to the solution of higher
acrylamide concentration to stabilize the gradient as
the solution is pumped in. Decrease the concentration
of initiator so that polymerization occurs from top to
bottom. This minimizes convective mixing due to the
heat generated by polymerization.
2
Close both the mixing port and outlet port, if
appropriate, on the gradient maker. Clamp the tubing
from the outlet when using the SG500.
3
Add initiator and catalyst, and immediately pour the
light (low concentration) acrylamide solution into the
mixing chamber—the chamber with the outlet port.
Open the mixing valve slightly to allow the connecting
channel to fill and force out air bubbles.
Close the valve again and pour the heavy (high concen-
tration) acrylamide solution into the reservoir chamber.
4
Start the magnetic stirrer and unclamp or open the
outlet valve. Start the pump and open the mixing valve.
Warning! Acrylamide is a
neurotoxin. Wear gloves
when handling acrylamide or
polyacrylamide. Wear a dust
mask when weighing acrylamide
or preparing acrylamide stocks.
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