Prevalence of E. coli ST131 among Uropathogenic E. coli Isolates from Iraqi Patients in Wasit Province, Iraq
Table 3
Components of 20 μl PCR master mix (2 pools) and amplification condition for detection of E. coli ST131 clone by RT-PCR assay.
Pool
Sterile double DW (μl)
Primers
DNA (μl)
Amplification conditions
ST131T
13
1 μl each of (i) ST131T.F (ii) ST131T.R
5
(1) Initial denaturation at 95°C for 5 min (2) 40 cycles of (i) Denaturation at 95°C for 5 s (ii) And annealing at 58°C for 10 s (iii) The fluorescence signal was measured at the extension step (iv) Following amplification, a melting curve was generated by heating the PCR product to 95°C with a ramp rate of 0.05°C/s
