Primer sequences for α-thalassemia multiplex PCR and expected amplicon sizes
| Name . | 5′→3′ sequence . | GenBank ID: nucleotides . | Concentration . | Amplicon (size) . |
|---|---|---|---|---|
| LIS1-F | ATACCATGGTTACCCCATTGAGC | HSLIS10:510→532 | 0.5 μM } | LIS1 3′UTR fragment (2350 bp) |
| LIS1-R | AGGGCTCATTACATGTGGACCC | HSLIS10:2859→2838 | 0.5 μM | |
| α2/3.7-F | CCCCTCGCCAAGTCCACCC | HUMHBA4:5676→5694 | 0.2 μM } | -α3.7jxna fragment (2022/2029 bp) |
| 3.7/20.5-R | AAAGCACTCTAGGGTCCAGCG | HUMHBA4:11514→11494 | 0.2 μM | |
| α2/3.7-F | As above | As above | — } | α2gene (1800 bp) |
| α2-R | AGACCAGGAAGGGCCGGTG | HUMHBA4:7475→7457 | 0.2 μM | |
| 4.2-F | GGTTTACCCATGTGGTGCCTC | HUMHBA4:3064→3084 | 0.5 μM } | -α4.2 jxn fragment (1628 bp) |
| 4.2-R | CCCGTTGGATCTTCTCATTTCCC | HUMHBA4:8942→8920 | 0.5 μM | |
| SEA-F | CGATCTGGGCTCTGTGTTCTC | HSGG1:26120→26140 | 0.2 μM } | --SEA jxn fragment (1349 bp) |
| SEA-R | AGCCCACGTTGTGTTCATGGC | HSCOS12:3817→3797 | 0.2 μM | |
| THAI-F | GACCATTCCTCAGCGTGGGTG | HSGG1:9592→9612 | 0.3 μM } | --THAI jxn fragment (1153 bp) |
| THAI-R | CAAGTGGGCTGAGCCCTTGAG | HSCOS12:1241→1221 | 0.3 μM | |
| 20.5-F | GCCCAACATCCGGAGTACATG | HSGG1:17904→17924 | 0.2 μM } | -(α)20.5 jxn fragment (1007 bp) |
| 3.7/20.5-R | As above | As above | — | |
| MED-F | TACCCTTTGCAAGCACACGTAC | HSGG1:23123→23144 | 0.2 μM } | --MED jxn fragment (807 bp) |
| MED-R | TCAATCTCCGACAGCTCCGAC | HSGG1:41203→41183 | 0.2 μM | |
| FIL-F | TTTAAATGGGCAAAACAGGCCAGG | HSGG1:12304→12327 | 1.0 μM } | --FIL jxn fragment (546 bp) |
| FIL-R | ATAACCTTTATCTGCCACATGTAGC | HSCOS12:570→546 | 1.0 μM |
| Name . | 5′→3′ sequence . | GenBank ID: nucleotides . | Concentration . | Amplicon (size) . |
|---|---|---|---|---|
| LIS1-F | ATACCATGGTTACCCCATTGAGC | HSLIS10:510→532 | 0.5 μM } | LIS1 3′UTR fragment (2350 bp) |
| LIS1-R | AGGGCTCATTACATGTGGACCC | HSLIS10:2859→2838 | 0.5 μM | |
| α2/3.7-F | CCCCTCGCCAAGTCCACCC | HUMHBA4:5676→5694 | 0.2 μM } | -α3.7jxna fragment (2022/2029 bp) |
| 3.7/20.5-R | AAAGCACTCTAGGGTCCAGCG | HUMHBA4:11514→11494 | 0.2 μM | |
| α2/3.7-F | As above | As above | — } | α2gene (1800 bp) |
| α2-R | AGACCAGGAAGGGCCGGTG | HUMHBA4:7475→7457 | 0.2 μM | |
| 4.2-F | GGTTTACCCATGTGGTGCCTC | HUMHBA4:3064→3084 | 0.5 μM } | -α4.2 jxn fragment (1628 bp) |
| 4.2-R | CCCGTTGGATCTTCTCATTTCCC | HUMHBA4:8942→8920 | 0.5 μM | |
| SEA-F | CGATCTGGGCTCTGTGTTCTC | HSGG1:26120→26140 | 0.2 μM } | --SEA jxn fragment (1349 bp) |
| SEA-R | AGCCCACGTTGTGTTCATGGC | HSCOS12:3817→3797 | 0.2 μM | |
| THAI-F | GACCATTCCTCAGCGTGGGTG | HSGG1:9592→9612 | 0.3 μM } | --THAI jxn fragment (1153 bp) |
| THAI-R | CAAGTGGGCTGAGCCCTTGAG | HSCOS12:1241→1221 | 0.3 μM | |
| 20.5-F | GCCCAACATCCGGAGTACATG | HSGG1:17904→17924 | 0.2 μM } | -(α)20.5 jxn fragment (1007 bp) |
| 3.7/20.5-R | As above | As above | — | |
| MED-F | TACCCTTTGCAAGCACACGTAC | HSGG1:23123→23144 | 0.2 μM } | --MED jxn fragment (807 bp) |
| MED-R | TCAATCTCCGACAGCTCCGAC | HSGG1:41203→41183 | 0.2 μM | |
| FIL-F | TTTAAATGGGCAAAACAGGCCAGG | HSGG1:12304→12327 | 1.0 μM } | --FIL jxn fragment (546 bp) |
| FIL-R | ATAACCTTTATCTGCCACATGTAGC | HSCOS12:570→546 | 1.0 μM |
jxn, junction.