Table 2.
ASAP PCR conditions. The thermalcycler parameters used to generate ASAP amplicons in successive rounds of PCR are presented.
| ASAP PCR conditions | ||
| 1. Touchdown PCR | Temperature (Centigrade) | Duration (Minutes) |
| Denaturation | 94 | 4 |
| 10 Cycles | ||
| Denaturation | 94 | 0:40 |
| Annealing | 55 – 0.5/cycle | 0:40 |
| Extension | 72 | 0:40 |
| 25 Cycles | ||
| Denaturation | 94 | 0:40 |
| Annealing | 50 | 0:40 |
| Extension | 72 | 0:40 |
| Final extension | 72 | 7 |
| 2. Touchdown PCR | ||
| Denaturation | 94 | 4 |
| 10 Cycles | ||
| Denaturation | 94 | 0:40 |
| Annealing | 52 – 0.5/cycle | 0:40 |
| Extension | 72 | 0:40 |
| 25 Cycles | ||
| Denaturation | 94 | 0:40 |
| Annealing | 47 | 0:40 |
| Extension | 72 | 0:40 |
| Final extension | 72 | 7 |
| 3. Touchdown – extension PCR | ||
| Denaturation | 94 | 4 |
| 10 Cycles | ||
| Denaturation | 94 | 0:40 |
| Annealing | 52 – 0.5/cycle | 0:40 |
| Extension | 72 | 1:00 + .05/cycle |
| 25 Cycles | ||
| Denaturation | 94 | 0:40 |
| Annealing | 47 | 0:40 |
| Extension | 72 | 1:30 + 0.05/cycle |
| Final extension | 72 | 10 |