Recipes
| Lower Gel Buffer (separating) | |
|---|---|
| Tris Base | 187 g |
| SDS | 4 g |
| Bring pH to 8.8 w/HCl | |
| Bring volume up to 1 L with dH2O, filter and store at 4 °C | |
| Upper Gel Buffer (stacking) | |
|---|---|
| Tris Base | 60.5 g |
| SDS | 4 g |
| Bring pH to 6.8 w/HCl | |
| Bring volume up to 1 L with dH2O, filter and store at 4 °C | |
| Running Buffer | |
|---|---|
| Tris Base | 60.6 g |
| Glycine | 288.2 g |
| SDS | 20 g |
| Bring volume up to 20 L with dH2O | |
| 5x Sample Buffer | |
|---|---|
| 2 M Tris, pH 6.8 | 1.58 mL |
| SDS | 1 g |
| 2-Mercaptoethanol | 2.5 mL |
| Glycerol | 5 mL (1.26 g/mL) |
| Bromophenol Blue | small pinch |
| Bring volume to 10 mL with dH2O | |
| Separating Gel Recipes (for Bio-Rad mini gel) | |||||
|---|---|---|---|---|---|
| 7.5% | 10% | 12.5% | 14% | 15% | |
| 30% Acrylamide/Bis (37.5:1) | 2.5 mL | 3.3 mL | 4.2 mL | 4.65 mL | 5.0 mL |
| dH2O | 5.0 mL | 4.2 mL | 3.3 mL | 2.85 mL | 2.5 mL |
| Lower Gel Buffer | 2.5 mL | 2.5 mL | 2.5 mL | 2.5 mL | 2.5 mL |
| 10% APS | 75 µL | 75 µL | 75 µL | 75 µL | 75 µL |
| Temed | 10 µL | 10 µL | 10 µL | 10 µL | 10 µL |
| Stacking Gel | |
|---|---|
| 30% Acrylamide/Bis (37.5:1) | 0.75 mL |
| dH2O | 3.0 mL |
| Upper Gel Buffer | 1.25 mL |
| 10% APS | 50 µL |
| Temed | 10 µL |
| Coomassie Stock | |
|---|---|
| Coomassie R-250 | 0.15 g |
| dH2O | 100 mL |
| Stain 1 (1 L) | |
|---|---|
| Acetic Acid | 100 mL |
| Isopropanol | 100 mL |
| Coomassie Stock | 20 mL |
| dH2O | 780 mL |
| Stain 2 (1 L) | |
|---|---|
| Acetic Acid | 100 mL |
| Coomassie Stock | 20 mL |
| dH2O | 880 mL |
| Destain (1 L) | |
|---|---|
| Acetic Acid | 100 mL |
| dH2O | 900 mL |
| Gel Soak (1 L) | |
|---|---|
| 95% Ethanol (treat as 100%) | 300 mL |
| Glycerol | 10 mL |
| dH2O | |
Media
| LB-Lennox (1 L) | |
|---|---|
| Tryptone | 10 g |
| Yeast Extract | 5 g |
| dH2O | Bring to 1 L |
| Nutrient Broth Soft Agar (200 mL) | |
|---|---|
| Nutrient Broth | 1.6 g |
| NaCl | 1.0 g |
| Agar | 1.3 g |
| dH2O | 200 mL |
| SOC (100 mL) | |
|---|---|
| Tryptone | 2 g |
| Yeast Extract | 0.5 g |
| NaCl | 50 mg |
| MgCl2 | 0.2 g (or 1 mL 1 M MgCl2) |
| KCl | 18.6 mg (or 0.25 mL 1 M KCl) |
| dH2O | 100 mL |
| Glucose (add after autoclaving) | 2 mL of 1 M Glucose (sterile filtered) |