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.