Table 3. Primers used for the multiplex PCR.
| Primer | Sequence 5′−3′ | Tm (°C) | Amplicon size (bp) | Target |
|---|---|---|---|---|
| Multiplex PCR targeting genes coding for chloramphenicol/florfenicol resistance | ||||
| MT-ctrl-chloram-F1 | GCTTACCTCAGATAATGAGTCGTC | 54,8 | 172 | Prophage 1 (Control) |
| MT-ctrl-chloram-R1 | GCCAATAAGAGCCCTACTCTTC | 55 | ||
| MT-cat-F1 | CTATTTTGACAATACGCCCTGC | 54,3 | 448 | cat |
| MT-cat-R1 | CTTCCCAAACGTAAATATCGGC | 54 | ||
| MT-floR-F1 | TTGAGCCTCTATATGGTGATGC | 54,4 | 632 | floR |
| MT-floR-R1 | GTTGTCACGATCATTACAAGCG | 54,3 | ||
| Multiplex PCR targeting genes coding for sulfonamide resistance | ||||
| MT-ctrl_sul-F1 | TTCATTTCGTCTTGGGTCTAGC | 54,8 | 175 | Prophage 1 (Control) |
| MT-ctrl_sul-R1 | GGACTACAGATCTACCATAATCCG | 54 | ||
| MT-sul1-F1 | GGGCTACCTGAACGATATCC | 54,7 | 550 | sul1 |
| MT-sul1-R1 | CTAGGCATGATCTAACCCTCG | 54,4 | ||
| MT-sul2-F1 | ATCATCTGCCAAACTCGTCG | 55,2 | 449 | sul2 |
| MT-sul2-R1 | TTCTTGCGGTTTCTTTCAGC | 53,9 | ||
| Multiplex PCR targeting genes coding for tetracycline resistance | ||||
| First multiplex PCR targeting genes coding for tetracycline resistance | ||||
| MT-ctrltet1-F1 | CCAGAATGACGAATTGAATGTCG | 54,3 | 175 | Prophage 1 (Control) |
| MT-ctrltet1-R1 | GGACCTCTTTACTCCAGTCG | 54,4 | ||
| MT-tetA(E)pAsa4-F1 | GATGTCACACCTGAGGAATCC | 55,1 | 351 | tetA(E) |
| MT-tetA(E)pAsa4-R1 | TCCGAATAAAACCCATAATGTTGC | 53,9 | ||
| MT-tetApSN54b-F1 | CAAGCAGGATGTAGCCTGTG | 55,9 | 526 | tetA |
| MT-tetApSN54b-R1 | ATTGCCGATATCACTGATGG | 52,4 | ||
| Second multiplex PCR targeting genes coding for tetracycline resistance | ||||
| MT-ctrltet2-F1 | ATTCATTTCGTCTTGGGTCTAGC | 55 | 176 | Prophage 1 (Control) |
| MT-ctrltet2-R1 | GGACTACAGATCTACCATAATCCG | 54 | ||
| MT-tetHpAB5S9b-F1 | ACGACTGTCTGATAAATACGGC | 54,6 | 326 | tetH |
| MT-tetHpAB5S9b-R1 | ATATCGAGTGTGAAATAGCGGC | 54,9 | ||
| MT-tetA(C)pRAS3-F1 | CTGTAGGCATAGGCTTGGTTAT | 54,4 | 629 | tetA(C) |
| MT-tetA(C)pRAS3-R1 | CTGTCCTACGAGTTGCATGATA | 54,1 | ||
| MT-tetGs62-F1 | GGTTCGCATCAAACCATTCG | 54,8 | 460 | tetA(G) |
| MT-tetGs62-R1 | GCTTAGATTGGTGAGGCTCG | 55,6 | ||