Bio-Rad Model 491 Prep Cell and Mini Prep Cell - Manual (Page 35)

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Section 8
SDS-PAGE Gel Preparation
8.1 Gel Recipes
Table 8 can be used to prepare both analytical and preparative Tris-HCl acry lamide gels. Sections 8.2–8.3
provide detailed formulas for calculating specific gel percentages and volumes. Acrylamide/bis stock solution
of 30% (37.5:1) are used. The amounts listed for the components in Table 8 are based on a total vol ume of
10 ml. Determine the total volume needed and multiply each component by the appro priate number. As long
as SDS is included in the sample buffer and the upper elec trophore sis running buffer it can be left out of the
gels during their preparation.
Table 8. Gel preparation reference.
(Acrylamide Monomer)
%T
ddi H2O,
ml
Gel buffer solution*,
ml
Acrylamide/bis Solution
30% stock (37.5:1), ml
4 6.15 2.50 1.33
5 5.80 2.50 1.67
6 5.45 2.50 2.00
7 5.15 2.50 2.33
8 4.80 2.50 2.67
9 4.45 2.50 3.00
10 4.15 2.50 3.33
11 3.80 2.50 3.67
12 3.45 2.50 4.00
13 3.15 2.50 4.33
14 2.80 2.50 4.67
15 2.45 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
Table 9. Catalyst preparation reference.
Catalyst
10% APS*
(µl)
TEMED*
(µl)
Analytical resolving gel 50 5
Stacking Gel 50 10
Preparative resolving gel 25 2.5
Stacking gel 50 10
**Note: Amounts are per 10 ml gel volume. Different amounts of catalyst are added for analytical and preparative gels. To make 10%
APS, dissolve 100 mg in 1 ml of deionized water.
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