SDS Polyacrylamide Gels
Resolving Gels (30:0.14 ratio Acrylamide-Bis low bis)
all volumes in mls (Note: Two Gel is 1.5X the amounts)
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.
| Type | One Gel 12% | One Gel 15% | One Gel 18% | Two Gels 12% | Two Gels 15% | Two Gels 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 | .4 | .4 | .4 | .6 | .6 | .6 | |
| 1% APS | 2 | 2 | 2 | 3 | 3 | 3 | |
| TEME | .04 | .04 | .04 | .06 | .06 | .06 |
Stacking Gels (5% Acrylamide 0.13% Bis)
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.
| Type | One Gel | Two Gel |
|---|---|---|
| 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 | .975 | 1.4625 |
| 10% SDS | .15 | .225 |
| 1% APS | .75 | 1.125 |
| TEMED | .015 | .0225 |