Table 2. Polymerase Chain Reaction primers used to detect haplotype variation across two noncoding cpDNA regions in wild Cyclopia intermedia populations.

Primers used for High Resolution Melt analysis are denoted in bold typeface and primers used to amplify the full cpDNA region are denoted in italic typeface. The primers used for unidirectional sequencing of PCR products for HRM cluster confirmation are underlined and italicised. The average number of length of PCR products amplified by the various primer combinations are given in base pairs (bp), followed by the primers nucleotide motif, and the annealing temperature used for PCR.

cpDNA region	Primer ID	PCR product size (bp)	Sequence (5′- >3′)	Tm (°C)
ndhA intron	ndhAx1	1100	GCYCAATCWATTAGTTATGAAATACC	55
	ndhAx2		GGTTGACGCCAMARATTCCA
	MLT_U1	350	AGGTACTTCTGAATTGATCTCATCC	59.0
	MLT_U2		GCAGTACTCCCCACAATTCCA
atpI-atpHintergenic spacer	atpI	1100	TATTTACAAGYGGTATTCAAGCT	50
	atpH		CCAAYCCAGCAGCAATAAC
	MLT_S1	200	TGGGGGTTTCAAAGCAAAGG	58.8
	MLT_S2		ATTACAGATGAAACGGAAGGGC
	MLT_S3	300	TTCCCGTTTCATTCATTCACATTCA	59.3
	MLT_S4		CCTTTGCTTTGAAACCCCCA