Table 1 The concentration of PCR components used in determining the antibiotic resistance genes
Target resistance gene | PCR components and concentration used | ||||||
|---|---|---|---|---|---|---|---|
10 × PCR buffer | MgCl2 (mM) | dNTP (mM) | Primer cons. (µM) | Taq polymerase (U) | Template DNA (µl) | ||
Forward | Reverse | ||||||
tet(A) | 1 X | 2.5 | 0.2 | 0.2 | 0.2 | 1.5 | 10 |
tet(B) | 1 X | 2.5 | 0.2 | 0.2 | 0.2 | 1.5 | 10 |
tet(C) | 1 X | 2.5 | 0.2 | 0.52 | 0.52 | 1.5 | 10 |
qnr(A) | 1 X | 1.5 | 0.2 | 0.1 | 0.1 | 1.5 | 3 |
qnr(B) | 1 X | 2.5 | 0.2 | 0.1 | 0.1 | 1.5 | 3 |
qnr(S) | 1 X | 1.5 | 0.2 | 0.1 | 0.1 | 1.5 | 3 |
strA/strB | 1 X | 2.5 | 0.16 | 1 | 1 | 1 | 5 |
aac(3)IV | 1 X | 3 | 0.2 | 0.2 | 0.2 | 1.5 | 5 |
aadB | 1 X | 2.5 | 0.08 | 0.12 | 0.12 | 1 | 5 |
aphA1 | Master mix (12.5 µl) | 0.16 | 0.16 | - | 5 | ||
aphA2 | 1 X | 2.5 | 0.08 | 0.1 | 0.1 | 1 | 5 |
sul1 | 1 X | 2.5 | 0.2 | 0.2 | 0.2 | 1.5 | 10 |
sul2 | 0.3 | 0.3 | |||||
sul3 | 0.2 | 0.2 | |||||