Polymerase chain reactions (PCR) primers for sequence-specific PCR
| Target region | Primer | Sequence (5’→3’) | Binding site | Annealing temp. | Extension time | |
|---|---|---|---|---|---|---|
| 5’ UTR to exon 1 | F | gaactaagtcccaagccccg | 5’ UTR | Yes | 58℃ | 30 s |
| Exon 2 | F | gcagatggtgtggtagtcc | Intron 1 | Yes | 58℃ | 30 s |
| R | ggggtccattccctctatg | Intron 2 | No | |||
| Exon 3 | F | taaggctagtgtcttttcagg | Intron 2 | Yes | 60℃ | 30 s |
| R | ccaggttcatctctaactc | Intron 3 | Yes | |||
| Exon 4 | F | ctctgaacaccagtctcg | Intron 3 | Yes | 58℃ | 30 s |
| R | ctgcccgattttctattagcctc | Intron 4 | Yes | |||
| Exon 5-6 (1st) | F | aggctaatagaaaatcgggcag | Intron 4 | Yes | 60℃ | 1 min |
| R | ttgaagccaataagagaatgca | Intron 6 | Yes | |||
| Exon 5 (2nd) | F | aaatgtggttagctggtatcag | Intron 4 | No | 58℃ | 30 s |
| R | tgtgtgctagtcctgttagacc | Intron 5 | No | |||
| Exon 6 (2nd) | F | gcagcaagatggtgttctct | Intron 5 | No | 58℃ | 30 s |
| R | ttgaagccaataagagaatgca | Intron 6 | Yes | |||
| Exon 7 | F | ccatttggtggcgccggat | Intron 6 | Yes | 60℃ | 30 s |
| R | ctgctctgtgtttgtgggg | Intron 7 | Yes | |||
| Exon 8 (1st) | F | ggttgccttgaaggagagaga | Intron 7 | Yes | 58℃ | 30 s |
| R | ctgggattacaggtgccc | Intron 8 | Yes | |||
| Exon 8 (2nd) | F | gcccttcctggcaatggca | Intron 7 | No | 58℃ | 30 s |
| R | cacacggcagggtgcgct | Intron 8 | No | |||
| Exon 9 | F | ggtccaggaatgacagggcg | Intron 8 | Yes | 60℃ | 30 s |
| R | caggcatgtgctaccacg | Intron 9 | Yes | |||
| Exon 10 | F | caagagatcaagccaaaatcagt | Intron 9 | Yes | 58℃ | 30 s |
| R | ggaatgccagccaccttgtaaa | 3’ UTR | Yes | |||
| Human growth hormon (internal control) | F | tgccttcccaaccattccctta | - | 58℃ | 30 s | |
| R | ccactcacggatttctgttgtgtttc | - |