Table 1.
Primers (5′ to 3′) and sequences to analyze DNA methylation quantification via pyrosequencing for candidate genes.
| Primer/sequence to analyze | Avy assay | CabpIAP assay | Igf2 assay | Igf2r assay |
|---|---|---|---|---|
| Strand | Chr2: 154761458 strand = reverse | Chr2: 154179960–154180168 strand = reverse | Chr7: 149839707–149839926 strand = reverse | Chr17: 12935154–12935313 strand = reverse |
| Forward PCR primer | ATTTTTAGGAAAAGAGAGTAAGAAGTAAG | ATTATTTTTTGATTGGTTGTAGTTTATGG | TTTTTTAATATGATATTTGGAGATAGTT | TGGTATAATTAGAATTATAGTTTAAT |
| Reverse PCR primer | Biotin-CTACAAAAACTCAAAAACTCA | Biotin-CACCAACATACAATTAACA | Biotin-CCACATAATTTAATTCACTAATAATTACTA | Biotin-AAAAAACTCAAAAAATTCC |
| Sequencing primer | TAGAATATAGGATGTTAG | TAGAATATAGGATGTTAG | AATATGATATTTGGCGATAGTT | ATAATTAGAATTATAGTTTA |
| Sequence to analyze | YGTTATTTTGTGAYGGYGAATGTGGGGGYGGTT | YGTTATTTTGTGAYGGYGAATGTGGGGGYGGTT | YGYGGGAYGTTTGYGTAGAGGTTTGTTTGTTTTTTTGYGTGTTYGTYGGGGTYGT | ATYGGAATYGTATTAAAATTTTTYGAATTTTTGGGTAGYG |
| Amplicon length (bp) | 294 | 209 | 220 | 127 |
| Temperature (°C) | 53 | 47 | 56 | 52 |
| Cycles (n) | 42 | 40 | 50 | 50 |
Avy: Viable yellow agouti; CabpIAP: CDK5 activator-binding protein intracisternal A particle element; Chr: Chromosome.