抄録
Genetic engineering techniques were used in order to generate standard curves for the quantitative PCR of fecal markers. PCR amplification products derived from sewage, feces of pig, cow and black–tailed seagull were cloned into a TOPO-TA plasmid vector. Recombinant plasmids were inserted into Escherichia coli competent cells and transformed cells were screened on LB agar medium. Colony PCR and electrophoresis were used to confirm the DNA sequence length of each marker. The plasmids were extracted from transformed E. coli cells to generate standard curve for the quantitative PCR assay. Four standard curves for the quantifications of fecal markers were constructed and the PCR amplification efficiencies calculated from the slope were from 96 to 105%.
