Lysis Buffer
|
|
Final Concentration |
Stock Concentration |
For 10 ml Mix |
| Tris pH8.8 | 50 mM | 1 M | 500 µl |
| EDTA pH 8.0 | 1 mM | 0.5 M | 20 µl |
| Tween | 0.5% | 10% | 500 µl |
| H2O | 9000 µl |
- Add Proteinase K before use.
- For tail piece 0.2 mm long or ear piece, add 50 ul buffer plus 1.5 ul of 20 mg/ml (30 ug) Proteinase K stock solution.
- Ten samples: 550 ul lysis buffer + 16.5 ul PK 20 mg/ml.
- You can use 0.5 ml PCR tubes for digestion and heat in a PCR machine.
Lysis
- 55°C, O/N , then heat inactive at 100°C for ten minutes. Cool down on bench top.
- For 20 ul PCR reaction, use 0.8 ul tail DNA.