应用PCR-DHPLC技术高通量快速检测转基因玉米

High-throughput and Rapid Detection of Genetically Modified Corn by PCR-DHPLC Technology

利用多重PCR结合DHPLC技术首次建立转基因玉米高通量快速检测方法。根据公布的转基因玉米外源基因序列设计并合成适合多重PCR的引物序列,以转基因玉米为模板,通过优化多重PCR和DHPLC的反应条件,建立一套同时快速筛选检测玉米中多种转基因成分的7重PCR-DHPLC检测方法。该方法的检测灵敏度为0.195 ng/μL,质粒检测灵敏度为1×103拷贝/μL。

Established a method to detect transgenic corn by using the technology of Multiplex PCR with DHPLC rapidly and high-through for the first time. Multiplex PCR primers were designed and synthesized based on published transgenic corn exogenous gene sequences. To transgenic corn as a template, and established a seventh PCR-DHPLC detection method to screening a variety of genetically modified in corn simultaneously and rapidly by optimizing the multiplex PCR and DHPLC reaction conditions. The detection sensitivity of the method is 0.195 ng/lμL, and the de tection sensitivity of plasmid is 1 x 103 copy/lμL.