Table 3. Polymerase Chain Reaction and High Resolution Melt condition used to screen haplotype variation in wild Cyclopia intermedia populations.

Primer annealing temperature (Tm) for the various primer combinations used in this study are provided in Table 2.

Process	Step	Temperature	Time	Number of cycles
PCR Amplification	Initial Denaturing	95 °C	2 min	1
	Denaturing	95 °C	10 s	40
	Annealing + Plate Read	Primer Tm	30 s
	Extension + Plate Read	72 °C	30 s
HRM Analysis	Heteroduplex Formation	95 °C	30 s	1
		60 °C	1 min	1
	HRM + Plate Read	65–95 °C (in 0.2 °C increments)	10 sec/step	1