Table 3.
Primer sequences and annealing temperatures (Ta) used during MS-HRM analysis, amplicon length, number of CpG sites for each gene, accession number, and amplified region (* DMR2: differentially methylated region = upstream promoter region involved in the regulation of transcript 2 of DNMT3A gene, details are provided in [16]).
| Gene | Primer Sequences | Ta | Amplicon Length | CpG Sites | Accession Number | Amplified Region |
|---|---|---|---|---|---|---|
| MTHFR | F 5′-TTTTAATTTTTGTTTGGAGGGTAGT-3′ | 54° | 155 bp | 7 | NM_005957.4 | from +30 to +184 bp |
| R 5′-AAAAAAACCACTTATCACCAAATTC-3′ | ||||||
| DNMT1 | F 5′-GGTATCGTGTTTATTTTTTAGTAA-3′ | 52° | 114 bp | 9 | NG_028016.3 | from −106 to +8 bp |
| R 5′-ACGAAACCAACCATACCCAA-3′ | ||||||
| DNMT3A | F 5′-GGTTTGGGTTTATTGTAGGAAGGTTATTAAGGT-3′ | 58° | 199 bp | 7 | NM_153759.3 | DMR2 * |
| R 5′-AATCCAAAACCCCCCTATCACGAAA-3′ | ||||||
| DNMT3B | F 5′-TGGTGTTGTGTGATTATAGTGG-3′ | 55° | 174 bp | 6 | NG_007290.1 | from −397 to −223 bp |
| R 5′-TCACCCTAAAAAATCAAAAACC-3′ |