PCR and sequencing primers used in pyrosequence analysis for the DNA-methylation assay
| . | Sequence . | CpG dinucleotides . |
|---|---|---|
| γ-globin | ||
| Set 1 | ||
| PCR primer1 | TTGATAAGGTAAATTTGATTAATAGTTTTA | na |
| PCR primer2 | CCTCCTCTATAAAATAACCCATAAC | na |
| Set 2 | ||
| PCR primer1 | GAGTATTTAGTGAGGTTAGGGG | na |
| PCR primer2 | Biotin-ATACTTCCTTTTATTCTTCATCCC | na |
| Sequencing primer | ATTTAGTGAGGTTAGGGG | −54 and −51 |
| Set 3 | ||
| PCR primer1 | Biotin-TTGGTTAGGGATGAAGAATAAAAG | na |
| PCR primer2 | CCTCCTCTATAAAATAACCCATAA | na |
| Sequencing primer1 | AAAACTTATTAATAACCTCA | +5 and +16 |
| Sequencing primer2 | TCCTCTATAAAATAACCCAT | +49 |
| β-globin | ||
| Set 4 | ||
| PCR primer1 | GTGTAATAAGAAAATTGGGAAAA | na |
| PCR primer2 | Biotin-ATATCTCTTAACCCCATACCATCA | na |
| Sequencing primer1 | TGTAATAAGAAAATTGGGAA | −306 |
| Sequencing primer2 | AGTTGTGATTTTAAATATTA | −265 |
| Set 5 | ||
| Forward | GGGTTGAGGGTTTGAAGTTTAATT | na |
| Reverse | Biotin-CCAACCCTAAAATATAACTCCACA | na |
| Sequencing primer | AGAAGAGTTAAGGATAGGTA | −125 |
| . | Sequence . | CpG dinucleotides . |
|---|---|---|
| γ-globin | ||
| Set 1 | ||
| PCR primer1 | TTGATAAGGTAAATTTGATTAATAGTTTTA | na |
| PCR primer2 | CCTCCTCTATAAAATAACCCATAAC | na |
| Set 2 | ||
| PCR primer1 | GAGTATTTAGTGAGGTTAGGGG | na |
| PCR primer2 | Biotin-ATACTTCCTTTTATTCTTCATCCC | na |
| Sequencing primer | ATTTAGTGAGGTTAGGGG | −54 and −51 |
| Set 3 | ||
| PCR primer1 | Biotin-TTGGTTAGGGATGAAGAATAAAAG | na |
| PCR primer2 | CCTCCTCTATAAAATAACCCATAA | na |
| Sequencing primer1 | AAAACTTATTAATAACCTCA | +5 and +16 |
| Sequencing primer2 | TCCTCTATAAAATAACCCAT | +49 |
| β-globin | ||
| Set 4 | ||
| PCR primer1 | GTGTAATAAGAAAATTGGGAAAA | na |
| PCR primer2 | Biotin-ATATCTCTTAACCCCATACCATCA | na |
| Sequencing primer1 | TGTAATAAGAAAATTGGGAA | −306 |
| Sequencing primer2 | AGTTGTGATTTTAAATATTA | −265 |
| Set 5 | ||
| Forward | GGGTTGAGGGTTTGAAGTTTAATT | na |
| Reverse | Biotin-CCAACCCTAAAATATAACTCCACA | na |
| Sequencing primer | AGAAGAGTTAAGGATAGGTA | −125 |
na indicates not applicable.