大豆种子DNA快速提取方法的改良及应用朱振雷.pdfVIP

第40 卷第10 期 东 北 农 业 大 学 学 报 40(10): 60~63 2009 年 10 月 Journal of Northeast Agricultural University Oct. 2009 大豆种子DNA 快速提取方法的改良及应用 朱振雷，束永俊，李 勇，柏 锡，才 华，纪 巍，朱延明* （东北农业大学生命科学学院，哈尔滨 150030） 摘 要：高质量的DNA 是进行基因克隆等分子生物学试验的前提条件。因大豆种子中蛋白质和油脂含量丰 富，利用传统方法提取DNA 质量很难达到试验要求。研究在SDS 提取液的基础上，通过添加表面活性剂NP-40 和Tween-20 ，优化出一种适合于大豆种子DNA 快速提取的方法。优化后的SDS 方法提取的DNA 质量较高， OD260/OD280 在1.809～1.916 之间，无蛋白质和RNA 污染。对该方法提取的DNA 进行了基因克隆和分子标记的试验 验证，结果表明，它们能够用于大豆基因克隆、分子标记。 关键词：大豆种子；DNA 提取；SDS ；基因克隆；分子标记 中图分类号：S565.1 文献标识码：A 文章编号：1005-9369 （2009）10－0060－04 Improvement and application of the rapid DNA extraction method from soybean seeds/ZHU Zhenlei, SHU Yongjun, LI Yong, BAI Xi, CAI Hua, JI Wei, ZHU Yanming (College of Life Sciences, Northeast Agricultural University, Harbin 150030, China) Abstract: High-quality DNA is an important base of molecular biology manipulation, such as gene cloning. There are lots of protein and fat in the soybean seeds, so the DNA extracted from them by using traditional methods is very difficult to achieve quality requirements of experiments. This study improved the SDS extraction buffer by adding NP-40 and Tween-20, and established a rapid DNA extraction method that is suitable for soybean seeds. The DNA extraction by the optimized SDS extraction method has high-quality, and the values of OD260/OD280 ranged from 1.809 to 1.916, that means non-protein and RNA pollution. The DNA extraction method was validated by gene cloning and genotyping, and the results showed that it could be used for soybean gene cloning and molecular markers. Key words: soybean seeds; DNA extraction; SDS; gene cloning; molecular marker