基于纤维素膜吸附的黄瓜基因组DNA快速提取方法  

A rapid extraction method of cucumber genomic DNA based on cellulose membrane adsorption

作　　者：薛强[1] 王燕飞 盛万里 陈艳[1] 金涌[1] 邹明强[1] XUE Qiang;WANG Yanfei;SHENGWanli;CHEN Yan;JIN Yong;ZOU Mingqiang(Chinese Academy of Inspection and Quarantine,Beijing 100123,China;Solid Waste and Chemicals Management Center,Ministry of Ecology and Environment of the People's Republic of China,Beijing 100029,China;Technical Center of Hohhot Customs District,Hohhot 010020,Inner Mongolia,China)

机构地区：[1]中国检验检疫科学研究院,北京100123 [2]生态环境部固体废物与化学品管理技术中心,北京100029 [3]呼和浩特海关技术中心,呼和浩特010020

出　　处：《中国瓜菜》2021年第10期39-43,共5页China Cucurbits And Vegetables

基　　金：中国检验检疫科学研究院基本科研业务费项目(2019JK030)。

摘　　要：为快速获取与鉴定黄瓜基因组DNA,建立一种基于纤维素膜吸附的黄瓜基因组DNA快速提取方法,以市售华北型黄瓜为材料,制备纤维素膜片,筛选黄瓜裂解试剂,优化提取反应条件,建立了包括黄瓜裂解、膜片吸附、膜片清洗、核酸洗脱等步骤的黄瓜基因组DNA提取方法,可以在5 min内从黄瓜果实中提取基因组DNA用于PCR扩增。黄瓜果实的表皮、表层果肉和中心果肉都可以实现基因组DNA提取和PCR扩增,最少可以从13 mg黄瓜果实中提取并检测到黄瓜管家基因Act1片断。利用膜吸附DNA提取技术实现了黄瓜基因组DNA的快速提取,为黄瓜基因研究与快速鉴定提供了一种简便方法。In order to quickly obtain and identify cucumber genomic DNA, a rapid extraction method of cucumber genome DNA based on cellulose membrane adsorption is established. After preparation of cellulose membrane, development of lysis reagents, optimization of extraction conditions, a method includes cucumber lysis, membrane adsorption,membrane cleaning and nucleic acid elution was established. Genomic DNA of cucumber fruit could be extracted in 5 min and directly used for PCR amplification. Genomic DNA could be extracted from cucumber fruit epithelium, surface flesh and central flesh. And house-keeping gene Act1 fragments can be amplified from the 13 mg of cucumber fruit. Cucumber genomic DNA could be extracted quickly by using membrane adsorption DNA technology. It provides an easy method to research and rapid identification of cucumber gene.

关 键 词：黄瓜 膜吸附 基因组DNA提取 

分 类 号：S642.2[农业科学—蔬菜学]