一种适于PCR扩增的植物基因组快速提取新方法.pdfVIP

农业生物技术学报，2010 年，第18 卷，第2 期，第394-399 页 Journal of Agricultural Biotechnology, 2010, Vol.18, No.2,394-399 技术创新 Updated Technology 一种适于PCR 扩增的植物基因组快速提取新方法 1 1 1,2* 2 2 1,2 李筱婷 陈卓君 许文涛 元延芳 王一南 黄昆仑 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 * 通讯作者, xuwentao1111@ 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600 bp 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21 min ，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。 关键词 DNA 提取, 碱裂/ 醇沉法, 中和, PCR A New Simple and Rapid Plant DNA Extraction Method for PCR Analysis Li Xiaoting 1 Chen Zhuojun 1 Xu Wentao 1,2* Yuan Yanfang 2 Wang Yinan 2 Huang Kunlun 1,2 1 College of Food Science and Nutritional Engineering, China Agricultural University, Beijing, 100083; 2 The Supervision, Inspection and Testing Center of Genetically Modified Orginisms Food Safety, Ministry of Agriculture, Beijing, 100083 * Corresponding author, xuwentao1111@ DOI: 10.3969/j.issn.1674-7968.2010.02.031 Abstract A new DNA extraction method was established based on the traditional alkaline lysis method. In com- parison with the former method, the conditions of the extraction reaction were optimized. The volume of neutral- ization solution was adjusted and the isopropanol precipitation step was added for the purification of DNA. The re- sult 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 min- utes, which largely shortened the extraction time. And the new method also avoided the use of toxic reagents such as tris-phenol. Keywords DNA extraction, Alkaline