Bio-Rad Whole Gel Eluter and Mini Whole Gel Eluter - Manual (Page 18)

E. 10x Electrode (Running) Buffer, pH 8.3 (Makes 1 Liter)
Tris base 30.3 g
Glycine 144.0 g
SDS 10.0 g
Dissolve and adjust to 1,000 ml with deionized water. DO NOT adjust pH with acid or
base. To make 1 liter of 1x electrophoresis buffer (0.025 M Tris, 0.192 M glycine, 0.1% SDS,
pH 8.3) dilute 100 ml of 10x stock with 900 ml of deionized water.
6.2 Preparing SDS-PAGE Gels of Any %T
The following table can be used to prepare Tris-HCl acrylamide gels from 4% to 17 % T.
Sections 6.3 and 6.4 provide detailed formulas for calculating specific gel types.
Acrylamide/Bis stock solution of 30% (37.5:1) is used. The amounts listed for the components
in the table below are based on a total volume of 10 ml. When SDS is included in the sample
buffer and the upper electrophoresis running buffer it can be left out of the gels during their
preparation.
% T Deionized Gel buffer Acrylamide/Bis solution
(acrylamide H
2
O solution* 30% stock (37.5:1)
monomer) (ml) (ml) (ml)
4% 6.15 2.50 1.33
5% 5.80 2.50 1.67
6% 5.55 2.50 2.00
7% 5.15 2.50 2.33
8% 4.80 2.50 2.67
9% 4.47 2.50 3.00
10% 4.17 2.50 3.33
11% 3.80 2.50 3.67
12% 3.47 2.50 4.00
13% 3.15 2.50 4.33
14% 2.80 2.50 4.67
15% 2.47 2.50 5.00
16% 2.15 2.50 5.33
17% 1.80 2.50 5.67
* Resolving Gel buffer - 1.5 M Tris-HCl, pH 8.8
* Stacking Gel buffer - 0.5 M Tris-HCl, pH 6.8
Catalysts 10% APS* TEMED*
Resolving Gel 50 µl 5 µl
Stacking Gel 50 µl 10 µl
* Amounts are per 10 ml gel volume. To make 10% APS, dissolve 100 mg in 1 ml of deionized water. TEMED is used neat.
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