Table 2. Primer extension set.
| Primer/product numbera | Primer/product nameb | Mutation interrogated | Primer sequence | Used at,µMc | Product size, ntd | N/M genotypee |
| 1 | IVS-I-1+ | IVS-I-1 (G->A) | GGTGAGGCCCTGGGCAG | 1.6 | 18 | G/A |
| 2 | Codon 8− | Codon 8 (-AA) | ACAGGGCAGTAACGGCAGAC | 2.4 | 21 | T/C |
| 3 | IVS-I-110− | IVS-I-110 (G->A) | CAGCCTAAGGGTGGGAAAATAGAC | 0.3 | 25 | C/T |
| 4 | IVS-I-110+ | IVS-I-110 (G->A) | ATAGGCACTGACTCTCTCTGCCTATT | 0.1 | 27 | G/A |
| 5 | IVS-I-1− | IVS-I-1 (G->A) | CTTAAACCTGTCTTGTAACCTTGATACCAA | 0.4 | 31 | C/T |
| 6 | Codon 39− | Codon 39 (C->T) | (C)9 ATCCCCAAAGGACTCAAAGAACCTCT | 0.06 | 36 | G/A |
| 7 | IVS-I-6+ | IVS-I-6 (T->C) | (C)18 AGGCCCTGGGCAGGTTGG | 2.4 | 37 | T/C |
| 8 | Codon 5+ | Codon 5 (-CT) | (C)15 CAGACACCATGGTGCATCTGACTC | 1.2 | 40 | C/G |
| 9 | Codon 6/S+ | Codon 6 (-A) | (C)17 ACACCATGGTGCATCTGACTCCTG | 0.8 | 42 | A/G |
| 10 | IVS-II-745+ | IVS-II-745 (C->G) | (C)16 CATATTGCTAATAGCAGCTACAATCCAG | 0.8 | 45 | C/G |
| 11 | Codon 6/S− | Codon 6 (-A) | (C)27 GCAGTAACGGCAGACTTCTCC | 0.8 | 49 | T/C |
| 12 | Codon 5− | Codon 5 (-CT) | (C)28 CAGTAACGGCAGACTTCTCCTC | 0.8 | 51 | A/G |
| 13 | Codon 39+ | Codon 39 (C->T) | (C)27 GCTGCTGGTGGTCTACCCTTGGACC | 0.4 | 53 | C/T |
| 14 | IVS-I-6− | IVS-I-6 (T->C) | (C)24 GTCTCCTTAAACCTGTCTTGTAACCTTGAT | 0.8 | 55 | A/G |
| 15 | IVS-II-745− | IVS-II-745 (C->G) | (C)26 TATCCCAACCATAAAATAAAAGCAGAATGGTA | 0.1 | 59 | G/C |
a
Primers are numbered according to the order of the corresponding normal peaks on the electropherogram (Figure 1B).
b
Name ending on “+” indicates that primer is identical to the coding strand of the gene (for reference, see HBB sequence in NCBI entry NG_000007.3); name ending on “−” signifies identity to the opposite, template strand.
c
Final concentration in primer extension reaction.
d
Length of extension product in nucleotides (nt) = primer length in nt +1 nt.
e
Added terminator for normal (N) and mutant (M) template.