利用微滴式数字PCR技术分析转基因玉米抗除草剂标记基因EPSP拷贝数

发布时间：2018-07-12 19:02

本文选题：微滴式数字PCR + 转基因玉米　；参考：《华北农学报》2017年03期

【摘要】：基于微滴式数字PCR(Droplet digital PCR,ddPCR)平台,以转基因玉米为例,建立了转基因作物(Genetically modified crops,GM crops)外源基因拷贝数分析方法,对待测样品进行了快速鉴定,并从T_0转基因玉米株系中鉴定出多个单拷贝单株。对该方法与实时荧光定量PCR(Quantitative real-time PCR,qRT-PCR)方法在分析结果的准确性方面进行了比较,从试验数据可以看出,2种检测方法的结果比较一致,单拷贝检测结果高度一致;但是ddPCR试验操作更加简便,试验结果可重复性强,试验数据更加准确可靠。研究表明,ddPCR方法是一种更加便捷、快速和准确的外源基因拷贝数分析新方法,基于其在准确性和灵敏度方面的显著优势,将会在转基因作物的外源基因拷贝数分析中得到广泛的应用。
[Abstract]:Based on droplet digital PCR (droplet digital PCR) platform and taking transgenic maize as an example, a copy number analysis method of exogenous genes in transgenic crops (genetically modified cropsp GM crops) was established, and the samples were identified quickly. Multiple single copy plants were identified from transgenic maize lines. The accuracy of this method was compared with that of quantitative real-time PCR (quantitative real-time PCR). From the experimental data, we can see that the results of the two methods are consistent, and the results of single copy detection are highly consistent. But the ddPCR test is more simple, the test results are more reproducible, and the test data are more accurate and reliable. Studies have shown that the PCR method is a more convenient, rapid and accurate method for the analysis of copy number of foreign genes, based on its remarkable advantages in accuracy and sensitivity. It will be widely used in foreign gene copy number analysis of transgenic crops.
【作者单位】：长江大学农学院主要粮食作物产业化湖北省协同创新中心;北京市农林科学院北京农业生物技术研究中心北京市农业基因资源和生物技术重点实验室;
【基金】：国家转基因重大专项重点课题(2014ZX0800303B) 北京市农林科学院科研能力创新项目北京市科委项目(Z171100001517001)
【分类号】：S513

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