Assay ID Primer Sequence CpG-SNP PCR forward primer 5’-AAGAGTTAGAGTAGGATTTTAAGTAGAAT-3’ PCR reverse primer 5’-CTAAAAAACTCTCCAAACCTAATTTC-3’* Pyroseq primer 5’-TAGTTGATGTGTTAGTTTTTTG-3’ CpG site P1 PCR forward primer 5’-AAATTAAAGGGAAATAGGTATAAATTGT-3’ PCR reverse primer 5’-TAACACCAACTAACTAACAATACCC-3’* Pyroseq primer 5’-GGGAAATAGGTATAAATTGTAT-3’ CpG sites P2 and P3 PCR forward primer 5’-AAATTAAAGGGAAATAGGTATAAATTGT-3’ PCR reverse primer 5’-TAACACCAACTAACTAACAATACCC-3’* Pyroseq primer 5’-ATAGTTGGTTTGGATTATGTTGT-3’ *All reverse primers were labeled with biotin.

Table 2: PyroMark assays designed for DNA methylation analysis of the IGF1 gene.