Table 1.

Genes and Primers for Bisulfite Pyrosequencing

Gene	Primer	Sequence	Amplicon length (bp)	Localization (bp)	Number of CpGs
H19	Outer forward	5′-TTTTTGGTAGGTATAGAGTT-3′
	Outer reverse	5′-AAACCATAACACTAAAACCC-3′
	Nested forward	5′-TGTATAGTATATGGGTATTTTTGGAGGTTT-3′		Chromosome 11
	Nested reverse*	5′-TCCTATAAATATCCTATTCCCAAATAACC-3′	231	1977,647–1977,878	4
	Sequencing	5′-TGGTTGTAGTTGTGGAAT-3′
MEG3	Forward	5′-GATTTTTTTTATATATTGTGTTTG-3′		Chromosome 14
	Reverse*	5′-CTCATTTCTCTAAAAATAATTAACC-3′	220	100,361,907–100,362,129	3
	Sequencing	5′-GTGTTTGAATTTATTTTGTTTGG-3′
LIT1	Forward	5′-AATTAGTAGGTGGGGGG-3′		Chromosome 11
	Reverse*	5′-CTAAAAAACTCCCTAAAAATC-3′	122	2677,751–2677,873	2
	Sequencing	5′-GGGGGTAGTYGGAG-3′
NESP55	Outer forward	5′-TTTTTTATTTTATAGGGTGTATTT-3′
	Outer reverse	5′-AAAATAAAATACTTAAACACCAC-3′
	Nested forward	5′-TTTTTGTAGAGTTAGAGGGTAGGT-3′		Chromosome 20
	Nested reverse*	5′-AAAAAAAACAACTCAAAATCTACC-3′	343	56,848,506–56,848,849	3
	Sequencing	5′-GTGTTTAAGAGGATGGAT-3′
PEG3	Forward	5′-GGTGTAGAAGTTTGGGTAGTTG-3′		Chromosome 19
	Reverse*	5′-CTCACCTCACCTCAATACTAC-3′	153	62,043,756–62,043,909	4
	Sequencing	5′-TGTTTATTTTGGGTTGGT-3′
SNRPN	Forward*	5′-AGGGAGTTGGGATTTTTGTATT-3′		Chromosome 15
	Reverse	5′-CCCAAACTATCTCTTAAAAAAAAC-3′	237	22,751,105–22,751,342	2 × 3 = 6
	Sequencing 1	5′-ACACAACTAACCTTACCC-3′			3
	Sequencing 2	5′-CCAACCTACCTCTAC-3′			3

This table provides the primer sequences for bisulfite pyrosequencing of the six studied genes, the length and chromosomal localization (ensembl 54, May 2009) of the amplified segments, and the number of CpG sites in the amplicons. Biotinylated primers are indicated by an asterisk.