Chelex-100快速提取用于转基因检测DNA模板的研究  

Study on Rapid Method for Extracting DNA from Transgenic Crops by Chelex-100

作　　者：王永[1] 张莉[2] 兰青阔[1] 赵新[1] 程奕[1] 

机构地区：[1]天津市农业科学院中心实验室,天津300384 [2]天津市畜牧兽医研究所,天津300112

出　　处：《生物技术通报》2008年第2期143-145,共3页Biotechnology Bulletin

基　　金：科技部国际合作项目(2006DFA32380)

摘　　要：目的:建立从转基因作物中快速提取DNA的方法。方法:采用Chelex-100法提取抗草甘膦大豆和非转基因大豆、转基因抗虫玉米Bt176和非转基因玉米中的DNA,使用PCR扩增大豆和玉米的内源基因(Lectin,zSSIIb)及外源特异性序列(CaMV35S,Bt176)评价提取核酸的质量。结果:Chelex-100法能够快速在1h之内从大豆和玉米中提取DNA,所提取的DNA可以直接用于PCR扩增反应,PCR扩增产物电泳条带清晰,转基因抗草甘膦大豆样品和转基因抗虫玉米Bt176检测均出现强阳性结果。结论:Chelex-100法提取DNA可以作为转基因检测的模板,该方法具有经济、简便、快速的特点,适合于转基因检测工作。Objective：To establish a rapid method for extracting DNA from transgenic crops.Methods：DNA was extracted DNA from transgenic roundup ready soybean and transgenic Bt176 Maize by Chelex-100.The quality of the extraction by endogenous gene （kectin and aSSIIb）and foreign specific sequence （CoMV35S and Bt176）will be evaluated with PCR technique.The PCR product of CoMV35S and Bt176 was used as a mark fragment, Results：The Chlex-100 can quickly extract the DNA from transgenic crops about 1 hour,The DNA can be directly applied to the following step-PCR amplification as DNA template. Electrophoresis results of the PCR products are very clear,and the detection results of transgenic roundup ready soybean and transgenic Bt176 Maize showed strong positive,Conclusions：Chelex-100 extraction method could be an efficient and reliable method to extract from transgenic crops as DNA template for PCR detection

关 键 词：Chelex-100 DNA提取PCR CnMV35S Bt176 

分 类 号：S188[农业科学—农业基础科学]