一种油菜及其近缘植物干种子基因组DNA提取方法  

A DNA Extraction Method from Rapeseed and Closely Related Plants Seeds

作　　者：范惠玲 白生文 侯丽娟 Fan Hui-ling;Bai Sheng-wen;Hou Li-juan(College of Agricultural and Ecological Engineering,Hexi Univeristy;College of Life Science and Engineering,Hexi University,Zhangye Gansu 734000)

机构地区：[1]河西学院农业与生态工程学院 [2]河西学院生命科学与工程学院,甘肃张掖734000

出　　处：《河西学院学报》2021年第2期54-59,共6页Journal of Hexi University

摘　　要：种子DNA提取是种子分子生物学研究最基础和最关键的步骤之一.为了建立一种能够从油菜及其近缘植物干种子中制备高丰度、高纯度DNA的方法,本试验分别以三大类型油菜(Brassica rape)、芸芥(Eruca sativa M.)和白芥(Sinapis alba L.)的干种子为材料,用一定量的抽提液来研磨种子,提取基因组DNA,并通过测定不同OD值和琼脂糖凝胶电泳分别检测DNA样品液的纯度、浓度和完整性.结果表明,DNA带型清晰,未发生降解;DNA样品OD260/OD280的值在1.792~1.905之间,均值为1.861,即纯度较高;OD320的值介于0.000~0.002之间,均值为0.0008,即DNA样品中悬浮物的量较少;所有DNA样品液的OD260/OD230的值均大于2.0,即DNA样品没有被碳水化合物、盐类所污染.PCR扩增产物的条带清晰可辨,能够实现多态性片段的分离.总之,本试验的材料易得,操作简便、实用,易于实施,在油菜种质资源分析、种子纯度鉴定乃至种子分子机理探索等方面具有重要意义.DNA extraction from the seed becomes the most important and the base step for research on seed mo⁃lecular biology.A rapid and effective genomic DNA preparation method was established in this study.Dry seeds of rapeseed,Eruca sativa M.,Sinapis alba L.were grined in CTAB(Cetyl Trimethyl Ammonium Bromide)solution,DNA was isolated and its quantity and quality were evaluated by nucleic acid and protein analyzer,integrity were de⁃tection by electrophoresis on agrose gel.Results showed that DNA bands in the agrose gel are clear,little degrada⁃tion;The OD260/OD280 ratio was 1.792~1.905,its average was 1.861.The value of OD320 was in the range of 0.000~0.002,little suspended solids;The OD260/OD230 ratio was high than 2.0,indicated that DNA samples were not contami⁃nated by carbohydrates and salts.The bands of PCR amplified products were clear and distinguishable,polymorphism fragments could be isolated.For its characters of simple operations,materials obtained easily,easy-implementation and affordable cost,this method is of great importance for analysis of germplasm resources,identification of genetic purity of seed and molecular marker assistant selection.

关 键 词：油菜 芸芥 白芥 干种子 DNA提取 

分 类 号：Q94[生物学—植物学]