|
Final gel concentration |
| Light solution |
5% |
7.5% |
10% |
12.5% |
15% |
| Monomer stock solution |
8.4 ml |
12.5 ml |
16.7 ml |
21.0 ml |
25 ml |
| 4X Resolving gel buffer |
12.5 ml |
12.5 ml |
12.5 ml |
12.5 ml |
12.5 ml |
| 10% SDS |
0.5 ml |
0.5 ml |
0.5 ml |
0.5 ml |
0.5 ml |
| H2O
|
28.5 ml |
24.4 ml |
20.2 ml |
16.0 ml |
12.0 ml |
| 10% Ammonium persulfate |
165 ml |
165 ml |
165 ml |
165 ml |
165 ml |
| TEMED |
16.5 ml |
16.5 ml |
16.5 ml |
16.5 ml |
16.5 ml |
| Total volume |
50 ml |
50 ml |
50 ml |
50 ml |
50 ml |
|
|
|
Final gel concentration |
| Heavy solution |
10% |
12.5% |
15% |
17.5% |
20% |
| Monomer stock solution |
16.7 ml |
21.0 ml |
25.0 ml |
29.2 ml |
33.3 ml |
| 4X Resolving gel buffer |
12.5 ml |
12.5 ml |
12.5 ml |
12.5 ml |
12.5 ml |
| Sucrose |
7.5 g |
7.5 g |
7.5 g |
7.5 g |
7.5 g |
| 10% SDS |
0.5 ml |
0.5 ml |
0.5 ml |
0.5 ml |
0.5 ml |
| H2O
|
16.2 ml |
11.7 ml |
7.7 ml |
3.5 ml |
0 ml |
| 10% Ammonium persulfate |
165 ml |
165 ml |
165 ml |
165 ml |
165 ml |
| TEMED |
16.5 ml |
16.5 ml |
16.5 ml |
16.5 ml |
16.5 ml |
| Total volume |
50 ml |
50 ml |
50 ml |
50 ml |
50 ml |
| If the gel will be subsequently stained using an ammoniacal silver staining method, include sodium thiosulfate in the gel
to reduce background levels of stain. Use 90 ml of 10% ammonium persulfate solution, 45 ml of 5% sodium thiosulfate, and 90
ml of TEMED per 50 ml of gel solution. Sodium thiosulfate slightly inhibits polymerization, thus the requirement for higher
amounts of TEMED. Ammonium persulfate must be added to the gel solution first, followed by sodium thiosulfate, and TEMED last.
|
