In this study, samples identification through detection of lacZ
and uidA genes using PCR simultaneously (multiplex PCR) was
approved. Primers listed in Table 1 are used to amplify 876 bp
and 147 bp fragments of lacZ and uidA genes, respectively.
Amplification reaction in a final volume of 25 mL contains 5 mL
of diluted bacteria or purified DNA; and 2.5 micro liters buffer
containing 10 Xmg, 1 mL dNTPs, 1.5 units of Taq enzyme and
2 mL of both primers were added to it. The PCR reaction steps
were as follows.
Initial denaturating temperature: 94 °C for 9 min and then 23
thermal cycles for amplification in PCR reaction as: 94 °C for 1
min, 55 °C for 1 min and 72 °C for 50 s and final amplification
cycle (extension) at 72 °C for 2 min. Corrected programs of PCR
are shown in Table 1.