Skip to main content

Advertisement

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   
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