环介导等温扩增法检测转基因玉米MON89034

A Specific Loop-mediated Isothermal Amplification Method for Detection of Transgenic Maize MON89034

摘要:

根据MON89034外源插入片段与植物基因组序列设计特异性引物，筛选最佳引物并对反应体系和反应条件进行优化，最终建立转基因玉米MON89034转化体特异性LAMP检测方法。对该方法进行了特异性、灵敏度、稳定性和重复性测试。结果表明：该方法能够特异性检测出MON89034玉米；检测其灵敏度达到1 pg；以转基因玉米MON89034 DNA标准品质量分数为1.00%，0.10%，0.05%的样品为模板，其稳定性好、重复性高，假阴性率为0。本试验设计的LAMP方法适用于特异性检测转基因玉米MON89034。

Abstract:

According to MON89034 exogenous insert sequences connection with the plant genome sequences, the best specific primers were designed, the reaction system and reaction conditions were optimized, and finally a loop-mediated isothermal amplification method for detecting transgenic maize MON89034 was established. Specificity, sensitivity, stability and repeatability of this method were tested. The results showed that the method can specifically detect maize MON89034, and the detection sensitivity of the method was up to 1pg.With the maize MON89034 DNA samples of 1.00%, 0.10%, 0.05% concentration as templates, stability and repeatability testing was conducted, false negative rate was 0. The results showed that the LAMP method is suitable for specifically detecting transgenic maize MON89034.