| 1. Washing buffer | 10–15 min | 50 °C |
| 2. Washing buffer | 30–60 min | 50 °C |
| 3. Washing buffer | 30–60 min | 50 °C |
| 4. NTE | 5 min | 37 °C with slowly shaking in incubator |
| 5. NTE | 5 min | 37 °C with slowly shaking in incubator |
| 6. RNase A | 30 min | 37 °C with slowly shaking in incubator |
| 7. NTE | 5 min | room temperature shaking |
| 8. NTE | 5 min | room temperature shaking |
| 9. Washing buffer | 30–60 min | 50 °C |
| 10. PBS | 5 min | room temperature shaking |