| TYPE OF GEL: DISCONTINUOUS SDS-PAGE USING OWL PENGUIN SYSTEM |
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| RESOLVING GEL: (variable %T, 0.375 M Tris-HCl, pH 8.8) |
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| Volumes are in mL unless noted; total volume is 10 mL, just enough for two mini-gels. You may choose to make 15 mL. |
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Final Acrylamide Percentage |
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| Stock Solution |
5 |
7.5 |
10 |
12.5 |
15 |
17.5 |
20 |
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| Acrylamide-bisacrylamide |
1.25 |
1.88 |
2.50 |
3.13 |
3.75 |
4.38 |
5.00 |
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| (40% T, 3.33% C) |
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| Distilled water |
6.10 |
5.47 |
4.85 |
4.22 |
3.60 |
2.97 |
2.35 |
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| 10% SDS |
0.1 |
0.1 |
0.1 |
0.1 |
0.1 |
0.1 |
0.1 |
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| 1.5 M Tris-HCl, pH 8.8 |
2.5 |
2.5 |
2.5 |
2.5 |
2.5 |
2.5 |
2.5 |
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| 10% (w/v) Ammonium persulfate |
50 µL |
50 µL |
50 µL |
50 µL |
50 µL |
50 µL |
50 µL |
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| (make fresh regularly) |
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| Temed |
5 µL |
5 µL |
5 µL |
5 µL |
5 µL |
5 µL |
5 µL |
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| (after degassing) |
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| STACKING GEL: (4%T, 0.125 M Tris-HCl, pH 6.8) |
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| Volumes are in mL unless noted; total volume is 10 mL enough for two mini-gels |
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| Stock Solution |
10 mL |
5 mL |
2.5 mL |
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| Acrylamide-bisacrylamide |
1 |
0.5 |
0.25 |
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| (40% T, 3.33% C) |
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| Distilled water |
6.35 |
3.17 |
1.59 |
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| 0.5 M Tris-HCl, pH 6.8 |
2.50 |
1.25 |
0.63 |
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| 10% SDS |
0.1 |
0.05 |
0.025 |
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| 10% (w/v) Ammonium persulfate |
50 µL |
25 µL |
12.5 µL |
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| Temed |
10 µL |
5 µL |
2.5 µL |
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