SDS Polyacrylamide Gels

all volumes in mls

Resolving Gels      (30:0.14 ratio Acrylamide-Bis low bis)

                                                        One Gel Two Gels (1.5X the amounts)        
12% 15% 18%         12% 15% 18%
ddH2O 14.44 11.16 7.88 21.66 16.74 11.82
1.5 M Tris, pH 8.6 10 10 10 15 15 15
40% Acrylamide solution 12 15 18 18 22.5 27
2% Bis solution 1.12 1.4 1.68 1.68 2.1 2.52
10% SDS 0.4 0.4 0.4 0.6 0.6 0.6
1% APS 2 2 2 3 3 3
TEMED 0.04 0.04 0.04 0.06 0.06 0.06

Pour. Overlay with water-saturated n-butanol (2-methyl propanol). The gel should polymerize in about 30 minutes.
Pour off alcohol, rinse 2X with distilled water . Dry top of gel thoroughly with filter paper inserted to the level of the gel.

Stacking Gels      (5% Acrylamide 0.13% Bis)

One Gel Two Gels
ddH2O 9.335 14.0025
1M Tris- HCl pH 6.8 1.9 2.85
40% Acrylamide Solution 1.875 2.8125
2% Bis Solution 0.975 1.4625
10% SDS 0.15 0.225
1% APS 0.75 1.125
TEMED 0.015 0.0225


Pour stacking gel between the glass plates to the very top.
Insert comb, allowing it to displace some of the liquid. (Remember acrylamide is a neurotoxin).
This should avoid bubbles forming. Let sit about 30 minutes or until polymerized.