Bio-Rad Mini-PROTEAN® Tetra Cell - Manual (Page 25)

21
Gel Formulations (10 ml)
1. Prepare the monomer solution by mixing all reagents except
the TEMED and 10% APS. Degas the mixture for 15 min.
30 % Degassed
Percent DDI H
2
O Acrylamide/Bis Gel buffer 10% w/v SDS
gel (ml) (ml) (ml) (ml)
4% 6.1 1.3 2.5 0.1
5% 5.7 1.7 2.5 0.1
6% 5.4 2.0 2.5 0.1
7% 5.1 2.3 2.5 0.1
8% 4.7 2.7 2.5 0.1
9% 4.4 3.0 2.5 0.1
10% 4.1 3.3 2.5 0.1
11% 3.7 3.7 2.5 0.1
12% 3.4 4.0 2.5 0.1
13% 3.1 4.3 2.5 0.1
14% 2.7 4.7 2.5 0.1
15% 2.4 5.0 2.5 0.1
16% 2.1 5.3 2.5 0.1
17% 1.7 5.7 2.5 0.1
* Resolving Gel Buffer – 1.5 M Tris-HCl, pH 8.8
* Stacking Gel Buffer – 0.5 M Tris-HCl, pH 6.8
2. Immediately prior to pouring the gel, add:
For 10 ml monomer solution:
Resolving gel: 50 µl 10% APS and 5 µl TEMED
Stacking gel: 50 µl 10% APS and 10 µl TEMED
Swirl gently to initiate polymerization.
Note: Prepare any desired volume of monomer solution by
using multiples of the 10 ml recipe. The volumes of APS and
TEMED must be adjusted accordingly.
Warning: The catalyst concentration is very important!
Webbing and incomplete well formulation can result from
inaccurate catalyst concentration.