Hoefer SE640 - Manual (Page 38)

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p30
The total percent of acrylamide (% T) in the
resolving gel, which can range from 5 to 20%,
determines the pore size. Commonly, the amount
of crosslinker used (% C) is 2.6%. In the fol-
lowing example system, the resolving gel com-
position is 10% T, 2.6% C, which results in a
medium pore size. The stacking gel composition
is 4% T, 2.6% C. The % T in the stacking gel is
lower because a larger pore size is required.
Final concentrations
Separating gel Stacking gel Electrophoresis buffer
Acrylamide conc. 10% T*, 2.6% C 4% T, 2.6% C
Tris-Cl 0.375 M 0.125 M
Tris-Glycine 0.025 M Tris base
0.192 M glycine
pH 8.8 6.8 ~8.3
SDS 0.1% 0.1% 0.1%
Ammonium persulfate (APS) 0.05% w/v 0.05 – 0.1% w/v
TEMED † 0.05% v/v 0.05 – 0.1% v/v
* To achieve any other desired final concentration, adjust the acrylamide stock and water volumes. Volumes for different
concentrations are listed on page 34.
† Tetramethylethylenediamine