Table 1 Protocol for multiple cycle fluorescence detection.
Step | Reaction | Temperature (ºC) | Time (s) | Number of cycles |
|---|---|---|---|---|
Activation | 95 | 30 | – | |
Amplification 1 | Denaturation | 93 | 1 | 7 |
Polymerisation | 64 | 1 | ||
Detection cycle 8 | Denaturation | 93 | 1 | 1 |
Polymerisation | 64 | 1 | ||
Amplification 2 | Denaturation | 93 | 1 | 6 |
Polymerisation | 64 | 1 | ||
Detection 15 | Denaturation | 93 | 1 | 1 |
Polymerisation | 64 | 1 | ||
Amplification 3 | Denaturation | 93 | 1 | 4 |
Polymerisation | 64 | 1 | ||
Detection cycle 20 | Denaturation | 93 | 1 | 1 |
Polymerisation | 64 | 1 | ||
Amplification 4 | Denaturation | 93 | 1 | 4 |
Polymerisation | 64 | 1 | ||
Detection cycle 25 | Denaturation | 93 | 1 | 1 |
Polymerisation | 64 | 1 | ||
Amplification 5 | Denaturation | 93 | 1 | 4 |
Polymerisation | 64 | 1 | ||
Detection cycle 30 | Denaturation | 93 | 1 | 1 |
Polymerisation | 64 | 1 | ||
Amplification 6 | Denaturation | 93 | 1 | 4 |
Polymerisation | 64 | 1 | ||
Detection cycle 35 | Denaturation | 93 | 1 | 1 |
Polymerisation | 64 | 1 |
