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运用实时荧光聚合酶链式反应技术(polymerase chain reaction,PCR)对转基因玉米MON863进行品种特异性检测和定量分析。通过设计玉米内源基因和外源基因边界序列特异性引物和Taqman-MGB探针,验证了内源基因的物种特异性和外源基因边界序列的品种特异性。利用已知转基因百分含量的MON863玉米作为标准品,进行荧光定量反应,建立定量标准曲线,通过标准曲线对玉米样品MON863玉米成份进行含量分析。结果表明,该方法重复性好,检测特异性强,最低检测限浓度达到0.001 ng/μL,即14个拷贝。由于使用实时荧光PCR技术,检测周期短,操作简便,可广泛运用于转基因玉米MON863的进出口检测和转基因产品的含量分析。
The MON863 transgenic corn was tested for its breed specificity by quantitative polymerase chain reaction (PCR). The species specificity of the endogenous genes and the variety specificity of the exogenous gene boundary sequences were verified by designing the primers specific for the endogenous maize and exogenous gene border sequences and the Taqman-MGB probe. The MON863 corn with known percentage of transgene was used as a standard to carry out fluorescence quantitative reaction, and a quantitative standard curve was established. The content of corn MON863 corn was analyzed by standard curve. The results showed that this method has good reproducibility and specificity, with the lowest detection limit of 0.001 ng / μL, ie 14 copies. Due to the use of real-time fluorescence PCR technology, the detection cycle is short, easy to operate, can be widely used in the import and export of transgenic corn MON863 import and export content analysis of genetically modified products.