TABLE 1.
The sequences and binding sites of primers used in this studya
| Primer name | Sequence (5′–3′) | Binding site |
|---|---|---|
| Forward | ||
| F1 | CGAAGTCAAAACTTTCACGCCCATTGAAAC | 1,521…1,550 |
| F2 | TTGACGCTTGGTTCATTGGCTTTACC | 1,688…1,713 |
| F3 | ATTACGGCACCATGCTCAAACCC | 1,466…1,488 |
| F4 | CTTTAAGCGACATGGGGTTTAAAAACCT | 1,358…1,385 |
| N562H.F | ATTGCCGGTAAAACCGGGACTTCTAACAACCATATTGACG | 1,654…1,693 |
| T558S.F | AACCGGGAGTTCTAACAACAATATTGACGCTTGGTTCATT | 1,665…1,704 |
| T593A.F2 | GGAGCGGCAGGAGGCGTTGTGAGCGCGCCTGT | 1,771…1,802 |
| G595S.F2 | ACCTATTGGCAAAGGAGCGACAGGAAGCGTTGTGAGCGC | 1,758…1,796 |
| Reverse | ||
| R1 | CGCTCCTTTGCCAATAGGTGTGT | 1,754…1,776 |
| R2 | GATGGAATTGGGAGTTGAATAGTAGGGGAT | 1,900…1,929 |
| R4 | GCGAAGGGTTGCATAAAATGTCTTTAGACG | 2,075…2,104 |
| N562H.R | CGTCAATATGGTTGTTAGAAGTCCCGGTTTTACCGGCAAT | 1,654…1,693 |
| T558S.R | AATGAACCAAGCGTCAATATTGTTGTTAGAACTCCCGGTT | 1,665…1,704 |
| T593A.R2 | CGCTCACAACGCCTCCTGCCGCTCCTTTGCCAATAGGT | 1,758…1,795 |
| G595S.R2 | GCGCTCACAACGCTTCCTGTCGCTCCTTTGCCAATAGGT | 1,758…1,795 |
a
The nucleotides underlined indicate the substitutions incorporated into the primer sequences. These modifications were specifically devised to swap the mutated nucleotide found in the pbp1 gene of the resistant strain KIN76 with the corresponding nucleotide from the susceptible strain 26695, in order to produce fused amplicons harboring each a single mutation.