User:Nader A. Khamis/Notebook/Experimental Biological Chemistry AU Fall 2011/2011/10/25
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The Stacking Gel, which has a lower concentration of acrylamide and pH compared to resolving gel, concentrates proteins into tight more easily visible bands.
The Resolving Gel seperates proteins according to their molecular weight.
The following manual was used to carry out the experimental procedure: 
1. Preparation of Stock Solutions:
A. 30% Acrylamide Mix, 10% SDS , and 1ml of 10 % Ammonium Persulfate (100 mg solid APS with 1 mL H2O)
B. Resolving Gel Solution (10 mL) : 3.3 mL H2O, 4.0 mL 30% Acrylamide mix, 2.5 mL Tris buffer (1.5M pH 8.8), 0.1 mL 10% SDS
C. Stacking Gel Solution (5 mL) : 3.4 mL H2O, 0.83 mL 30% Acrylamide mix, 0.63 mL Tris buffer (1.0M pH 6.8), 0.05 mL 10% SDS
A. Approximately 0.1 mL APS and 0.004 mL TEMED were pipetted to a 10 mL resolving gel solution
B. The gel plates was set up.
C. The gel comb was placed into plates and a mark was made 1 cm below the teeth using a permanent marker
D. The resolving gel solution was poured into plates until the mark.
E. Ethanol was added above the resolving gel solution until the top.
F. Allow gel to polymerize (20 minutes)
G. Excess Ethanol was removed.
H. Approximately 0.05 mL APS and 0.005 mL TEMED were pipetted to 5 mL stacking gel solution
I. The gel solution was poured into plates until it was full
J. The comb was inserted into the solution and gel was left for 20 minutes to polymerize.
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