Conditions for nested-PCR
| Amplification conditions | Temperature | Time | Cycle | |
|---|---|---|---|---|
| 1st PCR | Initial denaturation | 94°C | 5 min | 1 cycle |
| Denaturation | 94°C | 30 sec | 40 cycle | |
| Annealing | 58°C | 30 sec | ||
| Extension | 72°C | 40 sec | ||
| Final extension | 72°C | 5 min | 1 cycle | |
| 2nd PCR | Initial denaturation | 94°C | 5 min | 1 cycle |
| Denaturation | 94°C | 30 sec | 35 cycle | |
| Annealing | 58°C | 30 sec | ||
| Extension | 72°C | 40 sec | ||
| Final extension | 72°C | 5 min | 1 cycle | |
