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