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