一种适于PCR扩增的植物基因组快速提取新方法  

A New Simple and Rapid Plant DNA Extraction Method for PCR Analysis

作　　者：李筱婷[1] 陈卓君[1] 许文涛[1,2] 元延芳[2] 王一南[2] 黄昆仑[1,2] 

机构地区：[1]中国农业大学食品科学与营养工程学院,北京100083 [2]农业部转基因生物使用安全检验监督测试中心,北京100083

出　　处：《农业生物技术学报》2010年第2期394-399,共6页Journal of Agricultural Biotechnology

基　　金：国家高技术研究发展计划(863)项目(2006AA10Z440);国家自然基金(No.30800770);转基因生物重大专项(2008ZX08012-001)共同资助

摘　　要：在传统的碱裂解法提取基因组的基础上建立了一种新的基因组提取方法,这种方法对传统碱裂解制备基因组的方法进行优化,并且调整了中和剂的用量,引入异丙醇沉淀步骤对基因组进行纯化。实验证明这种方法扩大了碱裂解法制备基因组的适用范围,可直接从植物种子中快速提取出用于PCR(polymerase chain reaction)模板的基因组,对于600bp以下的目的基因的检测效果显著,可以发现待测组分含量大于1%的样品。整个基因组提取时间控制在21min,大大缩短了基因组提取时间,并且避免了酚/仿等对人体有害药品的使用。A new DNA extraction method was established based on the traditional alkaline lysis method.In comparison with the former method, the conditions of the extraction reaction were optimized.The volume of neutralization solution was adjusted and the isopropanol precipitation step was added for the purification of DNA.The result showed that the applicability of the new NaOH-based method was extended.DNA quickly extracted by this method could be directly used as temples to amplify fragments of the length of which were less than 600 bp.If the content is more than 1%, samples could be specificly amplified.The whole process could be finished in 21 minutes, which largely shortened the extraction time.And the new method also avoided the use of toxic reagents such as tris-phenol.

关 键 词：DNA提取 碱裂/醇沉法 中和 PCR 

分 类 号：Q943[生物学—植物学]