Real time PCR conditions and melt curve program for diagnosis of flaviviruses
| Cycle | Step | Temperature | Time |
|---|---|---|---|
| 1 cycle | Reverse transcription | 50°C | 15 min |
| Enzyme activation | 95°C | 10 min | |
| 40 cycles | Denaturation | 95°C | 10 sec |
| Annealing | 60°C | 15 sec | |
| Extension | 72°C | 10 sec | |
| Signal reading | 80°C | 30 sec | |
| 1 cycle | Denaturation | 95°C | 30 sec |
| 1 cycle | Melting |
60∼95°C | 15 sec |
| Cooling | 25°C | 30 min |
*Increase in temperature set at 0.5°C per cycle.
