Table 2.

Methylation specific PCR primer sequences

Gene	Nucleotide sequence	PCR product size, base pair (bp)
Methylation-independent primer sequences (nested)
RASSF1α	Forward: 5′-GGAGGGAAGGAAGGGTAAGG-3′	260
	Reverse: 5′-CAACTCAATAAACTCAAACTCCC-3′
p16 INK4a	Forward: 5′-GAGGAAGAAAGAGGAGGGGTTG-3′	274
	Reverse: 5′-ACAAACCCTCTACCCACCTAAATC-3′
Methylated allele-specific primer sequences
RASSF1α	Forward: 5′-GGGGGTTTTGCGAGAGCGC-3′	203
	Reverse: 5′-CCCGATTAAACCCGTACTTCG-3′
p16 INK4a	Forward: 5′-GAGGGTGGGGCGGATCGC-3′	143
	Reverse: 5′-GACCCCGAACCGCGACCG-3′
TIMP3	Forward: 5′-GCGTCGGAGGTTAAGGTTGTT-3′	116
	Reverse: 5′-CTCTCCAAAATTACCGTACGCG-3′
PCQAP 5′	Forward: 5′-GTTTTGTGATTGAGGYGGCGGC -3′	167
	Reverse: 5′-AAAAATCCCACAATCCAACCC -3′
PCQAP 3′	Forward: 5′-GATATGGGTGGTGGGAGTTGGG -3′	172
	Reverse: 5′- AATCAGACCCTAACCTCGCCCG -3′
Unmethylated allele-specific primer sequences
RASSF1α	Forward: 5′-GGTTTTGTGAGAGTGTGTTTAG-3′	172
	Reverse: 5′-ACACTAACAAACACAAACCAAAC-3′
p16 INK4a	Forward: 5′-TTATTAGAGGGTGGGGTGGATTGT-3′	145
	Reverse: 5′-CAACCCCAAACCACAACCATAA-3′
TIMP3	Forward: 5′-TGTGTTGGAGGTTAAGGTTGTTTT-3′	122
	Reverse: 5′-ACTCTCCAAAATTACCATACACACC-3′
PCQAP 5′	Forward: 5′-GTTTTGTGATTGAGGYGGTGGT -3′	167
	Reverse: 5′-AAAAATCCCACAATCCAACCC -3′
PCQAP 3′	Forward: 5′- TGATTAATTTAGATTGGGTTTAGAGAA -3′	158
	Reverse: 5′- CCAACTCCAAATCCCCTCTCTAT -3′