| Tris-HCl (pH 7.5) |
20 mm |
| KCl |
500 mm |
| MgCl2 |
2 mm |
| Puromycin |
1 mm |
| Dithiothreitol (DTT) |
1 mm |
| cOmplete protease inhibitor cocktail tablets, EDTA-free (Roche) |
2 tablets/100 mL culture |
| Heparin |
0.2 mg/mL |
| RNasin (Promega) |
1 µL/mL |
| Phenylmethylsulfonyl fluoride (PMSF) |
1 mm |
| Combine the first three ingredients. Just before use, add the puromycin and DTT, as well as the protease inhibitor tablets,
heparin, and RNasin (the latter three are optional but highly recommended). For processing meiotic cells, include PMSF.
|
