TABLE 1.

Thermal cycling conditions for SEFA PCR

Primer additionNo. of cyclesCycling conditions
Add 1 μl of 5 μM SP31(i) Denaturing at 94°C for 90 s; (ii) annealing at 35 to 45°C for 3 min and then ramping to 70°C at 0.2°C per s; and (iii) extension at 70°C for 5 min
Add 3 μl of 5 μM SP125(i) Denaturing at 94°C for 30 s and (ii) annealing and extension at 70°C for 5.5 min
Add no primers5 to 10 cycles of thermally asymmetric protocol(i) Two cycles of denaturing at 94°C for 30 s and annealing and extension at 70°C for 5 min; (ii) one cycle of denaturing at 94°C for 30 s, annealing at 55°C to 60°C for 30 s, and extension at 70°C for 5 min