1. Set the initial stage: corresponds topre-denaturation at 95°C for 10 minute.Only one cycle (single run)

Only one step

2. Set the cycling stage: corresponds toPCR amplification (38 cycles)

Two steps:

Step 1: 95°C for 15 seconds

Step 2: 60'C for 60 seconds(fluorescence signal is collected at thisstep)

The number of cycles for the cyclingstage should be set to 38, accordingto the manual.