Table 1

Optimum PCR concentrations and conditions

ParameterValue
Master mix PCR reagent23S rRNAparCgyrAgyrA2
    HotStarTaq master mix
    Forward primer (nM)200200200150
    Reverse primer (nM)200200200150
    MgCl2 (mM)1220
    DNA (10 μl)1:101:1001:101:10
Total vol (μl)25252525
PCR stepTemp (°C)TimeNo. of cycles
    Activation9510 min1
    Denaturation9510 s40
    Annealinga30 s40
    Extension7230 s40
    Final725 min1
  • a Annealing temperatures were 65°C for the 23S rRNA gene, 64°C for parC, 62°C for gyrA, and 61°C for gyrA2.