Step

Temperature

Time

Cycles

1

Reverse transcription

55 °C

15 min

1

2

Tag polymerase activation, pre-denaturing

95 °C

2 min

1

3

Denaturation

95 °C

15 sec

40

4

Annealing, extension, fluorescence acquisition

58 °C

35 sec

40

5

Instrument cooling

40 °C

10 sec

1