Table 2 MSP primers and PCR conditions.

Gene	Forward primer (5′–3′)	Reverse primer (5′–3′)	Tm and cycles	Size in bp	Locus	Reference
p15
M‐MSP	TGAGGATTTCG CGACGCGTTC	CGTACAATAA CCGAACGAC CGATCG	63°C/35	162	9p21	15
U‐MSP	TGAGGATTTTG TGATGTGTTT	CATACAATAA CCAAACAAC CAATCA		162
p16
M‐MSP	TTA TTA GAG GGT GGG GCG GAT CGC	GAC CCC GAA CCG CGA CCG TAA	65°C/35	150	9p21	15
U‐MSP	TTA TTA GAG GGT GGG GTG GAT TGT	CAA CCC CAA ACC ACA ACC ATA A		151
p18
M‐MSP	TTATCGAATTG TTATTTTCGTT CG	CGTCTCGCCG AAAAAATAA TC	64°C/35	93	1p32	15
U‐MSP	GGGTTATTGAA TTGTTATTTTT GTTTG	CATCCATCTC ACCAAAAAAA TAATC		95
RB
M‐MSP	GGGAGTTTCGC GGACGTGAC	ACGTCGAAAC ACGCCCCG	66°C/35	152	13q14	15
U‐MSP	GGGAGTTTTGT GGATGTGAT	ACATCAAAAC ACACCCCA		152
p21
M‐MSP	TTG GGC GCG GAT TCG TC	CTA AAC CGC CGA CCC GA	62°C/35	100	6p21	25
U‐MSP	TTA GTT TTT TGT GGA GTT G	CTC AAC TCT AAA CCA CAA		120
p27
M‐MSP	AAG AGG CGA GTT AGC GT	AAA ACG CCG CCG AAC GA	66°C/35	195	12p13	25
U‐MSP	ATG GAA GAG GTG AGT TAG T	AAA ACC CCA ATT AAA ACA		212
p57
M‐MSP	TCG GTT AGG TTT GAG CGA GC	TAC GTA TAC GAA AAA CGC GAC GAC	62°C/35	137	11p15	26
U‐MSP	TTG GTT AGG TTT GAG TGA GTG A	TCT ACA TAT ACA AAA AAC ACA ACA A		139		—

PCR, polymerase chain reaction; M, methylated; U, unmethylated; MSP, methylation‐specific PCR.