| Step | Temperature | Time | Cycles |
|---|---|---|---|
| Denaturation | 98 °C | 2 min | 1 |
| Denaturation | 98 °C | 10 s | 10 |
| Annealing | 72-62 °C (−1 °C/cycle) | 15 | 10 |
| Extension | 72 °C | 30 s | 10 |
| Denaturation | 98 °C | 10 s | 30 |
| Annealing | 62 °C | 15 s | 30 |
| Extension | 72 °C | 30 s | 30 |
| Hold | 4 °C | 1 | |
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