SDS Lysis Buffer
| Reagent | Amount to add (for 3.5 mL) | Final concentration (1×) |
|---|---|---|
| SDS (10%; Promega V6553) | 350 µL | 1% |
| Tris-HCl (1 m, pH 8.1) | 175 µL | 50 mm |
| EDTA (0.5 m, pH 8; Ambion AM9260G) | 70 µL | 10 mm |
| Complete Mini EDTA-free Proteinase Inhibitor Cocktail (Roche 11836170001) | 0.5 tablet | 1× |
| Phenylmethylsulfonyl fluoride (PMSF; 0.1 m in isopropanol; Fluka 93482) | 35 µL | 1 mm |
| Keep this buffer at room temperature to avoid SDS precipitation. Add protease inhibitors (Complete Mini EDTA-free Proteinase Inhibitor Cocktail and PMSF) immediately before using the buffer. | ||
