Primer sets for determination of human papillomavirus 16 CpG methylation using pyrosequencing
| Primer name | Primer sequence | Amplicon size (bp) |
|---|---|---|
| E2BS1 amplicon | ||
| Primary amplification forward primer Ampy E2BS1 F2 bio | 7348 5′-Biotin-ATTGTGTTGTGGTTATTTATTGTA-3′ 7371 | 175 |
| Primary amplification reverse primer Ampy E2BS1 R3 | 7522 5′-AACCATAATTACTAACATAAAACT-3′ 7499 | |
| Nested amplification forward primer Ampy E2BS1 F2 bio | 7348 5′-Biotin-ATTGTGTTGTGGTTATTTATTGTA-3′ 7371 | 142 |
| Nested amplification reverse primer Ampy E2BS1 R2 | 7489 5′-AACACATTTTATACCAAAAAAC-3′ 7468 | |
| Sequencing primer Sempy E2BS1 R2 | 7486 5′-ACATTTTATACCAAAAAACAT-3′ 7466 | – |
| E2BS2-4 amplicon | ||
| Primary amplification forward primer Ampy E2BS2,3,4 F | 7830 5′-TTGTAAAATTGTATATGGGTGTG-3′ 7852 | 193 |
| Primary amplification reverse primer Ampy E2BS2,3,4 R bio | 117 5′-Biotin-AAATCCTAAAACATTACAATTCTC-3′ 94 | |
| Nested amplification forward primer Ampy E2BS2 F | 7839 5′-TGTATATGGGTGTGTGTAAAT-3′ 7859 | 184 |
| Nested amplification reverse primer Ampy E2BS2,3,4 R bio | 117 5′-Biotin-AAATCCTAAAACATTACAATTCTC-3′ 94 | |
| Sequencing primer Sempy E2BS2 F | 7839 5′-TGTATATGGGTGTGTGTAAAT-3′ 7859 | – |
| Sequencing primer Sempy E2BS3,4 F | 9 5′-AATTTATGTATAAAATTAAGGG-3′ 30 | – |