Stage	Step	Condition
Hold	Step 1	Increase the temperature from 25 to 50°C with 1.6°C/s
Incubate at 50°C for 2:00 min
Step 2	Increase the temperature from 50 to 95°C with 1.6°C/s
Incubate at 95°C for 10:00 min
PCR	Step 1	Incubate at 95°C for 00:15 min
Step 2	Decrease the temperature from 95 to 60°C with 1.6°C/
Incubate at 60°C for 1:00 min
Number of cycles 40
Melt Curve	Step1	Increase the temperature from 60 to 95°C with 1.6°C/s
Step 2	Decrease the temperature from 95 to 60°C with 1.6°C/s
Incubate at 60°C for 1:00 min
Step 3 (dissociation)	Increase the temperature from 60 to 95°C with 0.05°C/s
Incubate at 95°C for 00:15 min