|
Final gel concentration |
|
5% |
7.5% |
10% |
12.5% |
15% |
| Monomer stock solution |
16.7 ml |
25 ml |
33.3 ml |
41.7 ml |
50 ml |
| 4X Resolving gel buffer |
25 ml |
25 ml |
25 ml |
25 ml |
25 ml |
| 10% SDS |
1 ml |
1 ml |
1 ml |
1 ml |
1 ml |
| H2O
|
56.8 ml |
48.5 ml |
40.2 ml |
31.8 ml |
23.5 ml |
| 10% Ammonium persulfate |
500 ml |
500 ml |
500 ml |
500 ml |
500 ml |
| TEMED |
33 ml |
33 ml |
33 ml |
33 ml |
33 ml |
| Total volume |
100 ml |
100 ml |
100 ml |
100 ml |
100 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 180 ml of 10% ammonium persulfate solution, 90 ml of 5% sodium thiosulfate, and
180 ml of TEMED per 100 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.
|
