Table 1.

List of Primers for MSP, RT-PCR, and Bisulfite Sequencing

Gene symbol	Analysis	Forward	Reverse	Tempera-ture (°C)	Number of cycles
ZIK1	MSP-M	5′-GTTTGAGGTGACGTTGGGC-3′	5′-GACCCTTTTCTCAACGCGA-3′	64	33
	MSP-U	5′-TTTGAGGTGATGTTGGGTG-3′	5′-AACAACCCTTTTCTCAACACA-3′	60	33
	RT-PCR	5′-AGAACTTTGCCCTTGTAGCC-3′	5′-GTGGATGAACCTGCCTTTG-3′	64	n/a
	Sequencing (X)	5′-AGGGTTTAGGTTAGAGTAGGAG-3′	5′-AAACTAAAACCAAATCCACAC-3′	52	30
	Sequencing (Y)	5′-AGTTTTTTGAGTTTGAATTAATG-3′	5′-ACAAAACCTTCTAAATTCTATAATC-3′	56	30
LOC-285300	MSP-M	5′-ATTTAAGCGGCGGTTCGT-3′	5′-CACGCGAAAAAACCAACG-3′	62	33
	MSP-U	5′-TTAAGTGGTGGTTTGTGGGTAT-3′	5′-AAAAAACAAAACCACCCACAC-3′	65	33
	RT-PCR	5′-CCGTTGCCGTGATAGTC-3′	5′-ACAGGAATCCGTGGTCAG-3′	60	n/a
CD8A	MSP-M	5′-GTTTGGTTAGGTTGTTTCGACG-3′	5′-ATCACCCGAAACCCCCG-3′	64	32
	MSP-U	5′-AGGTTGTTTTGATGGTTTTATTTATG-3′	5′-ATCACCCAAAACCCCCAC-3′	62	33
	RT-PCR	5′-CCGAGAGAACGAGGGCTAC-3′	5′-TCGCTGGCAGGAAGACC-3′	64	n/a
ZNF141	MSP-M	5′-CGGTTGTTATTTTCGTTATATTTC-3′	5′-ACTCCAATACGCTAATTAAATACG-3′	60	32
	MSP-U	5′-TACAACTCCAATCTCCCCA-3′	5′-TAGGTTAAGTATGAAGAGAAAGTTTT-3′	59	31
	RT-PCR	5′-CCTTTAGACGGTCCACAGAT-3′	5′-TATGTTGACTCAGGAGCGAA-3′	62	n/a
KAL1	MSP-M	5′-GTGCGAACGGGAGAGGC-3′	5′-GTCAACTACGAACCCGAACG-3′	64	33
	MSP-U	5′-AAAACCCATAAACCAATCTCA-3′	5′-TGAATGGGAGAGGTGTTTGT-3′	58	33
	RT-PCR	5′-TTGACAATGAGTGCTCTGGG-3′	5′-AATCGTAACTCTTTTCTGGGCT-3′	62	n/a
FGF14	MSP-M	5′-TTGTTTAGCGCGATTCGGATCGAC-3′	5′-CGAAAAATAAAACCGACCGACG-3′	60	33
	MSP-U	5′-TGTTTAGTGTGATTTGGATTGAT-3′	5′-ACAAAAAATAAAACCAACCAACA-3′	59	33
	RT-PCR	5′-GGCAACCTGGTGGATATCTT-3′	5′-CCTTGCCTGCAATATAACCT-3′	59	n/a
NKX2–8	MSP-M	5′-TTTTATTTTTTTCGGCGTTTTC-3′	5′-GAACGATAACCACCACCCG-3′	59	32
	MSP-U	5′-GTTTTTATTTTTTTTGGTGTTTTT-3′	5′-AAACAATAACCACCACCCA-3′	59	32
	RT-PCR	5′-AGCAGGACGCGCAACAC-3′	5′-AGTCTGGCGGGCTGGTC-3′	62	n/a
NID2	MSP-M	5′-GTTTTTAGAGTTAGGAGCGTTC-3′	5′-CGAATACGACTACTAACCTACG-3′	64	33
	MSP-U	5′-AGGTTAGTAGTTGTATTTGTTGTTGT-3′	5′-TCATCCAAACTACCCCACA-3′	58	33
	RT-PCR	5′-CTTACGAGGAGGTCAAACGC-3′	5′-CCGTTGGCAGGATAAAGAAA-3′	59	n/a
LOC-284801	MSP-M	5′-TGCGTTTACGTTCGTATTTGTC-3′	5′-GTAAAACCCGAAAAACCCG-3′	62	32
	MSP-U	5′-GTTGTGTTTATGTTTGTATTTGTT-3′	5′-CACCCCTCATAAAACCCA-3′	58	33
	RT-PCR	5′-GGCAATAAGAATGAAGACCT-3′	5′-ATCAACAGCCCTTAAACATC-3′	60	n/a
RAX	MSP-M	5′-AGTTGGTTTTTCGTTTTTAGCGTC-3′	5′-CTAAATCAAACTCCGCGAACG-3′	65	33
	MSP-U	5′-TAGTTGGTTTTTTGTTTTTAGTGTT-3′	5′-CCTAAATCAAACTCCACAAACA-3′	58	32
	RT-PCR	5′-TGCGTCTGAAAGCCAAGG-3′	5′-AGAGGATGCGGGTACGGT-3′	62	n/a
GAPDH	RT-PCR	5′-GCCAGTGGACTCCACGAC-3′	5′-CAACTACATGGTTTACATGTTC-3′	62	n/a

M, primers specific to methylated DNA; n/a, not available; U, primers specific to unmethylated DNA.