Table 2 Fast COLD-PCR Cycling Program
Step | Temp | Time | Analysis Mode | Acquisition Mode |
|---|---|---|---|---|
UNG | 40°C | 10 m | None | None |
Denaturation | 94°C | 10 m | None | None |
Regular-PCR (10 cycles) | 94°C | 1 s | None | None |
52°C | 20 s | None | None | |
72°C | 10 s | None | None | |
COLD-PCR (40 cycles) | 81 °C ( T c ) | 1 s | None | None |
Ramp 2.0°C/s | 52°C | 20 s | Quant | Single |
72°C | 10 s | None | None | |
Melting curve | 94°C | 5 s | None | None |
45°C | 30 s | None | None | |
80°C | Ramp 0.15°C/s | Melting | Continuous | |
Cooling | 40°C | 10 s | None | None |