Table 2.
The analysis stages of realtime PCR using LightCyler 2.0 system
| Analysis Stages |
Number of cycles |
Temperature (°c) | Duration | |
| Pre-incubation (To activate Taq DNA polymerase) |
1 | 95 | 10 minutes | |
| PCR cycle | 45 | Denaturation Annealing Extension |
95 58 72 |
10 seconds 10 seconds* 15 seconds |
| Melting analysis (to determine SNP) |
1 | Denaturation Annealing Melting |
95 40 85 |
0 second 30 seconds 0 second** |
| Cooling | 1 | 40 | 30 seconds | |
*
Single reading is taken to observe the amplification curve.
**
Continuous reading with 0.2 0C/sec ramp rate.