Primer sequences for Sanger sequencing
| Target region | Primer | Sequence (5’→3’) | Binding site | Amplicon size (bp) | |
|---|---|---|---|---|---|
| 5’ UTR to exon 1 | F | gaactaagtcccaagccccg | 5’ UTR (–836 to –817) |
Yes | 1146 |
| Exon 2 | F | tccagctgccatttagtaagactc | Intron 1 (–92 to –69) | No | 330 |
| R | ggggtccattccctctatg | Intron 2 (33 to 51) | No | ||
| Exon 3 | F | taaggctagtgtcttttcagg | Intron 2 (–703 to –683) | Yes | 1164 |
| R | ccaggttcatctctaactc | Intron 3 (292 to 310) | Yes | ||
| Exon 4 | F | ctctgaacaccagtctcg | Intron 3 (–132 to –115) | Yes | 701 |
| R | ctgcccgattttctattagcctc | Intron 4 (399 to 421) | Yes | ||
| Exon 5 | F | aaatgtggttagctggtatcag | Intron 4 (–177 to –156) | No | 398 |
| R | tgtgtgctagtcctgttagacc | Intron 5 (33 to 54) | No | ||
| Exon 6 | F | gcagcaagatggtgttctct | Intron 5 (–61 to –42) | No | 327 |
| R | ttgaagccaataagagaatgca | Intron 6 (107 to 128) | Yes | ||
| Exon 7 | F | ccatttggtggcgccggat | Intron 6 (–97 to –79) | Yes | 505 |
| R | ctgctctgtgtttgtgggg | Intron 7 (256 to 274) | Yes | ||
| Exon 8 | F | gcccttcctggcaatggca | Intron 7 (–103 to –85) | No | 257 |
| R | cacacggcagggtgcgct | Intron 8 (57 to 74) | No | ||
| Exon 9 | F | ggtccaggaatgacagggcg | Intron 8 (–164 to –145) | Yes | 826 |
| R | caggcatgtgctaccacg | Intron 9 (571 to 588) | Yes | ||
| Exon 10 | F | caagagatcaagccaaaatcagt | Intron 9 (–67 to 45) | Yes | 545 |
| R | ggaatgccagccaccttgtaaa | 3’ UTR (152 to 173) | Yes |
*Numbers of the primer binding sites indicate their distances from the adjacent exon/intron boundaries according to the reference sequences (accession No. NM_020485.8 and NG_0009208.3).
**The distances from the start codon are –875 to –856.