Hoefer SE640 Manual do Utilizador descarregar pdf (Página 43)

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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

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