Table 1

MSP and bisulphite sequencing primers for the SDHD, SDHB, and VHL promoters

Gene	Type of primer	Primer sequence (5′–3′)	Untreated DNA (5′–3′)	Annealing temperature
Y, C+T; R, G+A; U, primers for unmethylated DNA; M, primers for methylated DNA.
Primer sequences are given as forward and reverse for each set.
MSP
SDHB	U	TAAATGGGTATGTGTTGTTATTGT	CAAATGGGCATGCGCCGCTACTGC	60°C
		AACCACATACAAACAACCAAATAA	AGCCGCGTACGAGCAACCAAGTGG
	M	TGGGTATGCGTCGTTATTGC	TGGGCATGCGCCGCTACTGC
		CGCGTACGAACAACCAAATAA	CGCGTACGAGCAACCAAGTGG
SDHD	U	TGATAGTTGTGTTTGTGTATGTGT	CGACAGCTGTGTTTGCGCATGCGC	60°C
		AAACCACCATCTCATTCCTAAA	GAACCGCCATCTCGTTCCTGAG
	M	AGTTGTGTTTGCGTATGCGC	AGCTGTGTTTGCGCATGCGC
		AACCGCCATCTCGTTCCTAA	AACCGCCATCTCGTTCCTGA
VHL	U	GTTGGAGGATTTTTTTGTGTATGT	GCTGGAGGATCCTTCTGCGCACGC	60°C
		CCCAAACCAAACACCACAAA	CCCGGATCCCGCGGCGTCC
	M	TGGAGGATTTTTTTGCGTACGC	TGGAGGATCCTTCTGCGCACGC
		GAACCGAACGCCGCGAA	CGGATCCCGCGGCGTCC
Bisulphite
SDHB		GGAATAAAGAGGTTGAAATYGGG	GGAATAAAGAGGTTGAAATCGGGG	60°C
SDHB		AACCTACATCCACTAAAAACCCAC	AGCCTACATCCACTGAGGACCCAC	60°C
SDHD	1st set	TGAGGGGAATGGGATGTAGT	TGAGGGGAATGGGATGCAGC	60°C
		CTAAAAACTCAAAATCATCCACCAA	CTGAGGGCTCAAGGTCATCCACCAA	60°C
	2nd set	AGGGGATGGAAGTGAGGATTTA	AGGGGATGGAAGTGAGGACTCA	59°C
		AACAAACTCCTAACCCTCTTTC	GGCAAGCTCCTAACCCTCTTTC	59°C
VHL	1st set	TAGTGGAAATATAGTAAYGAGTTGG	TAGTGGAAATATAGTAACGAGTTGG	57°C
		CCCTCCRAAACATTCCCTC	CCCTCCGAAACATTCCCTC	57°C
	2nd set	TAYGGTGTTGGAGGATTTTTTTG	TACGGTGCTGGAGGATCCTTCTG	57°C
		TCTTCTTCAAAACCRTACTCTTC	TCTTCTTCAGGGCCGTACTCTTC	57°C