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Reaction conditions for SYBR Green-based Q-RT PCR analysis
| Name | Number of cycles | Step | Target temperature (°C) | Hold time | Ramp rate (°C/s) | Notes |
|---|---|---|---|---|---|---|
| Preincubation | 1 | 1 | 95 | 5 min | 4.4 | |
| Amplification | 45 | 1 | 95 | 10 s | 4.4 | |
| 2 | 60 | 10 s | 2.2 | |||
| 3 | 72 | 10 s | 4.4 | Fluorescence acquisition | ||
| Melt curve | 1 | 1 | 95 | 5 s | 4.4 | |
| 2 | 65 | 1 min | 2.2 | |||
| 3 | 97 | N/A | 0.11 | Continuous fluorescence acquisition (5/°C) | ||
| Cool | 1 | 1 | 40 | 30 s | 2.2 |
