Hoefer SE215 - Manual (Page 11)

Linear gradient gels
Gradient gels are prepared by first pumping the
“light” (lower % acrylamide) solutions and then
gradually adding a “heavy” (higher % acryl-
amide) into the caster by way of the inlet port
on the faceplate.
1
Follow instructions for preparing the gel caster on
page 2. Remove only the triangular rubber plug from
the bottom of the casting chamber, leaving the two
smaller plugs in place.
2
Follow the instructions for constructing the gel sand-
wich stack and loading it into the caster on page 3.
3
Connect the gradient maker to the gel caster. Run
a length of clear vinyl tubing through a peristaltic
pump and attach one end of the tubing to the gradi-
ent maker outlet port and the other end to the casting
chamber inlet port on the faceplate. Adjust the pump
to run a rate of about 5 ml/min for 4-gel casters and
about 10 ml/min for 10-gel casters.
•
p9
Fig 5. Gradient maker set-up.
Attach tubing to inlet port of the
multiple gel caster, run the tubing
through a peristaltic pump, and
attach to gradient maker.
Multiple gel caster
Peristaltic pump Magnetic stirrer
Mixing chamber
Gradient maker