Table 1 PCR primers used for bisulfite sequencing and methylation specific PCR

Bisulfite sequencing

Region1 Forward

5'-TAGTTTTTTGTTTAAAGGGTTTTAACGGGT-3'

365 bp

Region1 Reverse

5'- ACCTAAACTTAATAACTCCCCCTCA -3'

Region2 Forward

5'- ATTTTGGTATATTTTTGGGGAGTTT-3'

360 bp

Region2 Reverse

5'- CCCAAACACAAAATCTCATTTCTA -3'

Methylation specific PCR

Methylated Forward

5'-ATTTTTCGTCGGGAGTGGTC-3'

79 bp

Methylated Reverse

5'-GCAACGAATACGACGAACCT-3'

Unmethylated Forward

5'-GGGAGTGGTTGTGTGGTTTT-3'

103 bp

Unmethylated Reverse

5'-AACTTGGGGACACCCAAA-3'