Hoefer SE640 - Manual (Page 43)

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p35
For linear gradient gels, use equal volumes of
low % and high % acrylamide solutions in the
2 chambers of the gradient maker. Less APS is
added to extend polymerization time, and less
still is added to the higher %T solution to allow
polymerization to occur from the top down. In
our experience with the concentrations in the
10 – 20% gradient example below, gels can be
poured at a flow rate of 5 – 10 ml/min.
Linear gradient gel
(per 100 ml of solution)
10% T 20% T
Acrylamide stock (Solution #1) 33.30 ml 66.70 ml
Sucrose — 15.00 g
1.5 M TrisCl, pH 8.8 (Soln, #2) 25.00 ml 25.00 ml
10% SDS (Solution #4) 1.00 ml 1.00 ml
Deionized H2O a 100.00 ml a 100.00 ml
10% APS (Solution #5) 0.300 ml 0.060 ml
TEMED 0.036 ml 0.036 ml