| PCR cycling conditions | |||
|---|---|---|---|
| Steps | Temperature | Time | Cycles |
| Initial Denaturation | 95°C | 3 min | 1 |
| Denaturation | 98°C | 20 s | 13–15 cycles |
| Annealing | 65°C | 15 s | |
| Extension | 72°C | 1 min | |
| Final extension | 72°C | 5 min | 1 |
| Hold | 4°C | forever | |
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