基因组DNA快速提取及在转基因大豆检测中的应用  

Rapid extraction of genomic DNA and its application in detection of genetically modified soybeans

作　　者：陈欲 汪小福[2] 陈笑芸[2] 彭城 徐俊锋[2] 李玥莹[1] CHEN Yu;WANG Xiao-fu;CHEN Xiao-yun;PENG Cheng;XU Jun-feng;LI Yue-ying(College of Life Science,Shenyang Normal University,Shenyang 110034,China;State Key Laboratory for Man-aging Biotic and Chemical Threats to the Quality and Safety of Agro-products,Zhejiang Academy of Agricultural Sci-ences,Hangzhou 310021,China)

机构地区：[1]沈阳师范大学生命科学学院,辽宁沈阳110034 [2]农产品质量安全危害因子与风险防控国家重点实验室,浙江省农业科学院,浙江杭州310021

出　　处：《中国油料作物学报》2021年第1期70-76,共7页Chinese Journal of Oil Crop Sciences

基　　金：国家自然科学基金(31772098)。

摘　　要：基因组DNA的提取是DNA分子水平研究和检测的重要环节。为补充完善现场检测方法,根据硅膜吸附DNA的特性,结合过滤膜和注射器,开发一种现场快速提取植物基因组DNA的方法。选取大豆、棉花、油菜、玉米、水稻5种主要作物的叶片和种子为样品提取DNA,利用PCR和普通重组酶聚合酶扩增(recombinase polymerase amplification,RPA)对快速提取和QIAGNE试剂盒提取的DNA进行内源基因的扩增。结果显示,快速方法不需要离心机,现场3分钟完成DNA提取;在DNA得率上要高于QIAGEN方法,但在DNA纯度上低于QIAGEN方法。不过两种方法提取的DNA都能满足PCR和RPA的需要。利用DNA快速提取方法结合普通PCR、荧光RPA和荧光定量PCR对转基因大豆SHZD32-1进行实际检测,3种方法的检测结果一致,均能准确检出大豆SHZD32-1的转基因成分。To explore an on-site extraction method for plant genomic DNA during detection of genetically modified(GMO)soybeans,a rapid extraction of plant DNA was developed by applying silicon membrane to absorb DNA,together with filter membrane and syringe.The method was used to extract DNA from leaves and seeds of 5 major crops,including soybean,cotton,rapeseed,corn,and rice.The conventional PCR and basic RPA(recombinase polymerase amplification)are simultaneously used to amplify endogenous genes from the DNA extracted by the rapid extraction and QIAGEN kit methods.Results showed that the rapid extraction method had higher DNA yield than the QIAGEN method,while both of the extraction met the needs of conventional PCR and basic RPA amplification.By detecting the genetically modified soybean SHZD32-1,the DNA rapid extraction method combined with conventional PCR,fluorescence RPA and fluorescence quantitative PCR showed consistent and accurately results.For this developed rapid DNA extraction method does not require centrifuge and other equipment,and could be completed in 3 minutes on site,it was considered to be successfully meeting the requirements on site.

关 键 词：基因组 DNA快速提取 转基因检测 现场检测 

分 类 号：S565.1[农业科学—作物学] Q943.2[生物学—植物学]