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Touchdown PCR thermocycling conditions
| Initial denaturation | 94°C | 2 min | ||
| Touchdown | 11 cycles | 94°C | 30 sec | |
| 55°C | 30 sec | −1°C /cycle | ||
| 68°C | 2 min | |||
| Denaturation | 40 cycles | 94°C | 30 sec | |
| Annealing | 4x8°C | 30 sec | ||
| Extension | 68°C | 2 min | ||
| Final extension | 68°C | 10 min | ||
| Holding | 12°C | forever |
All the PCR conditions in this study are touchdown PCR, with adjustments to the annealing temperatures, and extension times.
