改良Chelex-100法快速提取转基因农产品DNA  

A Quick Method for DNA Extraction from Genetically Modified Agricultural Products by Improved Chelex-100

作　　者：蔡翠霞[1] 肖维威[1] 康文杰[1] 周琳华[1] 吴永彬[1] 郑远金[1] 马文丽[1] 

机构地区：[1]南方医科大学基因工程研究所,广州510515

出　　处：《生物技术通报》2011年第10期210-214,共5页Biotechnology Bulletin

基　　金：国家公益性行业科研专项(200910265);广东省科技计划项目(2008B080701027)

摘　　要：旨在建立一种从转基因农产品中快速提取DNA的方法。分别采用改良Chelex-100法和常规CTAB法提取转基因大豆GTS40-3-2基因组DNA,测其浓度和纯度,PCR扩增其内源基因(Lectin)、启动子(CaMV35S)和品系特异性序列,对两种方法进行比较和评价,并研究两种方法提取的DNA在-20℃下保存一个月内的检测效果,以及改良Chelex-100法在玉米、小麦和水稻等其他转基因农产品的应用效果。结果表明,改良Chelex-100法能够快速在1.5 h之内从样品中提取DNA,所提取的DNA直接用于PCR扩增反应,产物电泳条带清晰明亮。两种方法提取的DNA在-20℃下保存一个月内的检测效果未见明显差别。该方法在玉米、小麦和水稻等转基因农产品的应用效果稳定。因此,改良Chelex-100法提取的DNA可以作为PCR扩增模板用于转基因农产品检测。该方法具有经济、简便、快速、安全的特点,适合转基因农产品大规模筛选和鉴别。To establish a rapid method for extracting DNA from transgenic farm produce,we applied both improved Chelex-100 method and conventional CTAB method to extract total DNA from genetically modified soya.We compared the two methods by evaluating the quantity and purity of the extracted DNA detected by ultraviolet spectrometer and PCR amplification result of the endogenous gene（Lectin）、 promoter（CaMV35S） and variety specific sequence.The results indicated that the improved Chelex-100 method can quickly extract the DNA from samples within 1.5 hours,it can be directly applied to the following step-PCR amplification as DNA template.There is no significant difference between the PCR amplification of DNA extracted by the two methods and stored at-20℃ within 1 month.In addition,the application of the improved Chelex-100 method in maize,rice,wheat and other genetically modified agricultural products are stable.Therefore,the improved Chelex-100 extraction method could be an economic,simple and rapid method for PCR detection.It is suitable for large-scale screening and identification of genetically modified agricultural products.

关 键 词：Chelex-100法 CTAB法 DNA提取 PCR 

分 类 号：S188[农业科学—农业基础科学]