SDS Polyacrylamide

SDS Polyacrylamide Gels

 


all volumes in mls

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

 

(Note: Two Gel is 1.5X the amounts)

 One Gel 12%One Gel 15%One Gel 18% Two Gels 12%Two Gels 15%Two Gels 18%
ddH2O14.4411.167.8821.6616.7411.82
1.5 M Tris, pH 8.6101010151515
40% Acrylamide solution1215181822.527
2% Bis solution1.121.41.681.682.12.52
10% SDS.4.4.4.6.6.6
1% APS222333
TEME.04.04.04.06.06.06

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 GelTwo Gel
ddH2O9.33514.0025
1M Tris- HCl pH 6.81.92.85
40% Acrylamide Solution1.8752.8125
2% Bis Solution.9751.4625
10% SDS.15.225
1% APS.751.125
TEMED.015.0225


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.