Table 4.

Real-time PCR + melting conditions

Gene	F2/F5/12 (single, dúplex or triplex)
Cycles	Temperature (°C)			Time (seconds)	Ramp rate (°C/s)
1	95			180	20
45–50	95			15	20
	55			15	20
	72			30	20
Melting	95			60	20
	55			30	20
	45			30	20
	40			120	20
	85			0	0.1
Cool	40			30
Gene	MTHFR		H63D		C282Y
Cycles	Temp (°C)	Time (seconds)	Temp (°C)	Time (seconds)	Temp (°C)	Time (seconds)	Ramp rate (°C/s)
1	95	180	95	180	95	180	20
45–50	95	15	95	15	95	15	20
	53	15	60	15	55	15	20
	72	30	72	30	72	30	20
Melting	95	2	95	0	95	5	20
	40	20	45	0	45	15	20
	80	0	85	0	85	0	0.1
Cool	40	40	40	40	40	40

Abbreviation: PCR, polymerase chain reaction.