| Step | Temperature | Time |
|---|---|---|
| 1. Initial Denaturation | 98°C | 2 minutes |
| 2. Denaturation | 94°C | 15 seconds |
| 3. Annealing | 55°C | 15 seconds |
| 4. Extension | 72°C | 1 minute/kb |
| 5. Repeat steps 2–4 | 25 total cycles | |
| 6. Final extension | 72°C | 10 minutes |
| 7. Hold | 4°C | ∞ |
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