TABLE 4.

PCR conditions

Gene or regionsInitial denaturationAmplification (no. of cycles)aFinal hold
actA94°C, 2 min 30 s94°C, 3 min; 53°C, 1 min; 72°C, 2 min (40)72°C, 5 min
inlA94°C, 2 min 30 s94°C, 3 min; 66°C, 1 min; 72°C, 2.5 min (35)72°C, 5 min
sigB94°C, 5 min94°C, 30 s; TD 54-44°C, 30 s; 72°C, 1 min (40)72°C, 7 min
prs94°C, 5 min94°C, 30 s; TD 60-50°C, 30 s; 72°C, 1 min (40)72°C, 7 min
recA94°C, 2 min 30 s94°C, 30 s; 50°C, 30 s; 72°C, 1 min (40)72°C, 7 min
hly-mpl and plcA-hly94°C, 2 min 30 s94°C, 30 s; 54°C, 30 s; 72°C, 1 min (40)72°C, 7 min
  • a TD, touchdown PCR. For sigB and prs, a touchdown PCR protocol was used with annealing temperatures decreasing at a rate of 0.5°C/cycle from an annealing temperature T for the first 20 cycles followed by another 20 cycles with the annealing temperature set at T − 10°C.