Yeast Disruption Buffer
| Reagent | Final concentration |
|---|---|
| Tris-HCl (pH 7.4) | 50 mm |
| NaCl | 40 mm |
| Dithiothreitol (DTT) | 5 mm |
| MgCl2 | 5 mm |
| Glycerol | 5% |
| Benzonase (e.g., Sigma-Aldrich #E1014, 250 units/μL) | 0.5 µL/mL |
| RNase A | 0.1 mg/mL |
| Complete protease inhibitors with EDTA (Roche) | 1× |
| Phenylmethylsulfonyl fluoride (PMSF) | 5 mm |
| Use buffer immediately, or prepare and keep it without enzymes and DTT at 4°C for several weeks. Add the enzymes and DTT immediately before use. | |
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