Samples
were kept at 95 8C for 20 s (hold step) to allow DNApolymerase
activation and then subjected to 40 cycles
consisting of a denaturation step at 95 8C for 3 s followed
by an annealing/extension step at 60 8C for 30 s. Fluorescence
due to SYBR1 Green-1 incorporation was acquired at
the end of the extension step. No-template (NTC) and no-
RT controls were included in each experiment. A melting
curve analysis for specific amplification control was
performed (from 60 8C to 95 8C) at the end of the
amplification cycles. NTC controls were assumed as