At first, the reaction mixture was incubated at 96 C for 4 min. Then performed
35 amplification cycles at 94 C for 45s, 55 C for 60s, and 72 C for
60s. Reaction was further incubated for 7 min at 72 C. A positive
control (Escherichia coli genomic DNA) and a negative control were
included in the PCR. The PCR product was purified by using
Montage PCR Clean up kit (Millipore) in order to remove unincorporated
PCR primers and dNTPs from PCR products.