Reaction step

Temp (°C)

Time (sec)

Cycles

Analysis mode

Acquisition mode

Hot start

95

60

1

None

None

Amplification

95

5

55

Quantification

None

59

5

None

72

10

Single

Melting curves

55 to 95

20°C/sec (Ramp)

Melting curves

Continuous

Cooling

To 40

30

1

None

None

Table 2: Reaction condition of relative quantification of RT-PCR analysis.