| Step | Temperature | Time | Cycle number |
|---|---|---|---|
| Pre-incubation | 94 °C | 5 min | 1 |
| Denature | 94 °C | 30 s | Repeat 39 times |
| Annealing | 58 °C (adjusted by the GC content of the primers) | 30 s | |
| Extension | 72 °C | 40 s (product ~600 bp) | |
| Final extension | 72 °C | 5 min | 1 |
| Hold | 4 °C | - | 1 |
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