Step | Temperature (°C) | Time (min) |
---|---|---|
1. Denaturation | 95 | 5 |
2. Incubation | 60 | 25 |
3. Denaturation | 95 | 5 |
4. Incubation | 60 | 85 |
5. Denaturation | 95 | 5 |
6. Incubation | 60 | 175 |
Repeat step 1–6 one more time (two cycles of step 1–6 in total) | - | - |
7. Hold | 20 | Indefinite |