
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 |