| PCR cycling conditions | |||
|---|---|---|---|
| Steps | Temperature | Time | Cycles |
| Hold | 50°C | 5 min | 1 |
| Initial denaturation | 95°C | 5 min | 1 |
| Denaturation | 95°C | 10 s | 45 cycles |
| Annealing | 63°C | 15 s | |
| Extension | 72°C | 20 s | |
| Melting | Ramping from 65°C to 95°C | 1 | |
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