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目的:建立转Bt基因稻米的降落PCR-毛细管电泳快速定性检测方法。方法:转Bt基因稻米经磨碎后提取基因组DNA,对其内源蔗糖磷酸合酶SPS基因和抗虫毒蛋白Bt基因同时进行聚合酶链反应(PCR)扩增,产物用无胶筛分毛细管电泳分离、激光诱导荧光检测器检测。结果:在优化条件下,25 min内即可完成SPS和Bt基因的检测,日内精密度在2.37%～3.19%之间,特异性实验结果表明本法特异性较高。结论:所建立的方法快速、分离效能高、样品用量少且成本低廉,适用于转Bt基因稻米的快速鉴定。
Objective: To establish a rapid and qualitative PCR-capillary zone electrophoresis method for the detection of Bt transgenic rice. Methods: Genomic DNA was extracted from the transgenic Bt rice after milling. The endogenous sucrose phosphate synthase SPS gene and the anti-toxin protein Bt gene were simultaneously amplified by polymerase chain reaction (PCR) Electrophoresis separation, laser-induced fluorescence detector detection. Results: Under optimized conditions, the detection of SPS and Bt genes could be completed within 25 min. The intra-day precision was between 2.37% and 3.19%. The specific experimental results showed that the method was highly specific. Conclusion: The established method is rapid, high separation efficiency, less sample and low cost, and is suitable for the rapid identification of Bt transgenic rice.