Table 2 The target genes in the study, oligonucleotide primers, and PCR conditions used.
Target gene | Primers sequences (5`- 3`) | Product size (bp) | Primary denaturation | Number of cycles (No.) and PCR conditions | Final extension | References | |||
|---|---|---|---|---|---|---|---|---|---|
No. | Denaturation | Annealing | Extension | ||||||
aac(6’)/ aph(2’’) | F-GAAGTACGCAGAAGAGA | 491 | 94 ˚C 5 min. | 35 | 94 ˚C 30 s. | 54 ˚C 40 s. | 72 ˚C 40 s. | 72 ˚C 10 min. | |
R-ACATGGCAAGCTCTAGGA | |||||||||
ermB | F-CATTTAACGACGAAACTGGC | 405 | 94 ˚C 3 min. | 35 | 94 ˚C 30 s. | 55 ˚C 30 s. | 72 ˚C 1 min. | 72 ˚C 5 min. | |
R-GGAACATCTGTGGTATGGCG | |||||||||
tetM | F-GTGGACAAAGGTACAACGAG | 406 | 95 ˚C 5 min. | 30 | 95 ˚C 1 min. | 62 ˚C 1 min. | 72 ˚C 1 min. | 72 ˚C 7 min. | |
R-CGGTAAAGTTCGTCACACAC | |||||||||
vanA | F-CATGACGTATCGGTAAAATC | 885 | 95 ˚C 10 min. | 36 | 94 ˚C 1 min. | 56 ˚C 1 min. | 74 ˚C 1 min. | 74 ˚C 10 min. | |
R-ACCGGGCAGRGTATTGAC | |||||||||
int1 | F-CCTCCCGCACGATGATC | 280 | 96 ˚C 5 min. | 30 | 96 ˚C 30 s. | 55 ˚C 30 s. | 72 ˚C 30 s. | 72 ˚C 7 min. | |
R-TCCACGCATCGTCAGGC | |||||||||
int2 | F-TTGCGAGTATCCATAACCTG | 288 | 94 ˚C 5 min. | 35 | 94 ˚C 30 s. | 55 ˚C 30 s. | 72 ˚C 1 min. | 72 ˚C 7 min. | |
R-TTACCTGCACTGGATTAAGC | |||||||||
cylA | F-ACTCGGGGATTGATAGGC | 688 | 95 ˚C 15 min. | 30 | 94 ˚C 1 min. | 56 ˚C 1 min. | 72 ˚C 1 min. | 72 ˚C 10 min. | |
R-GCTGCTAAAGCTGCGCTT | |||||||||
gelE | F-TATGACAATGCTTTTTGGGAT | 213 | 95 ˚C 15 min. | 30 | 94 ˚C 1 min. | 56 ˚C 1 min. | 72 ˚C 1 min. | 72 ˚C 10 min. | |
R-AGATGCACCCGAAATAATATA | |||||||||