|
Separating Gel |
| |
12% GEL |
7.5% GEL |
| |
2 gels |
4 gels |
2 gels |
4 gels |
|
Milli Q water |
4.13 mL |
8.25 |
5.82 |
11.64 |
|
1.5M Tris-Cl pH 8.8 |
3.0 mL |
6.0 |
3.0 |
6.0 |
|
30% Acrylamide / Bis-Acrylamide(29:1) |
4.8 mL |
9.6 |
3.0 |
6.0 |
|
10% Ammonium Persulfate (fresh) |
60 uL |
120 |
60 |
120 |
|
TEMED |
6 uL |
12 |
6 |
12 |
| |
|
Stacking Gel |
| |
2 gels |
4 gels |
|
|
|
Milli Q water |
3.05 mL |
6.10 |
|
|
|
0.5 M Tris-Cl pH 6.8 |
1.25 mL |
2.50 |
|
|
|
30% Acrylamide / Bis-Acrylamide(29:1) |
0.67 mL |
1.33 |
|
|
|
10% Ammonium Persulfate (fresh) |
25 uL |
50 uL |
|
|
|
TEMED |
5 uL |
10 uL |
|
|
| |
|
|
|
|
|
Running Buffer (10X) |
|
Tris Base |
29g |
|
|
|
|
Glycine |
144 g |
|
|
|
|
Milli Q water |
To 1 Liter |
|
|
|
| |
|
|
|
|
|
Loading Buffer (2X) |
|
3.0 M Tris-Cl pH 8.45 |
167 uL |
|
|
|
|
0.1% Bromophenol Blue |
500 uL |
|
|
|
|
Glycerol |
2.0 mL |
|
|
|
|
Milli Q water |
To 10 mL |
|
|
|
| |
|
|
|
|
| |
|
|
|
|
Run gels at constant 125V for 1-12 hr.
Stain with filtered Brilliant Blue in 45% Methanol / 10% Glacial Acetic Acid
Destain with 45% Methanol / 10% Glacial Acetic Acid
