“…Program II contained no incubation at 42 °C and included shortened denaturation (5 s), annealing (15 s), and elongation (5 s) steps. Recently, it was shown that even shorter durations of these steps (1, 5 and 1 s, respectively) are sufficient for the formation of short DNA amplicons with Taq DNA polymerase [ 34 ]. Program III contained an RT step and had reduced denaturation temperature (85 °C), which is suitable for efficient amplification with nearby primers as well.…”