一种快速高效提取花生叶片DNA的简化方法  

A Simplified Method for Rapid Extraction of DNA from Peanut Leaves

作　　者：刘宇[1,2] 李春娟[2] 闫彩霞[2] 赵小波[2] 孔青[1] 孙全喜[2] 苑翠玲[2] 王娟 单世华[1,2] LIU Yu;LI Chun-juan;YAN Cai-xia;ZHAO Xiao-bo;KONG Qing;SUN Quan-xi;YUAN Cui-ling;WANG Juan;SHAN Shi-hua(College of Food Science and Engineering,Ocean Univ.of China,Qingdao 266000,China;Shandong Peanut Research Institute,Qingdao 266100,China)

机构地区：[1]中国海洋大学食品科学与工程学院,山东青岛266000 [2]山东省花生研究所,山东青岛266100

出　　处：《花生学报》2019年第1期58-61,共4页Journal of Peanut Science

基　　金：中央引导地方科技发展专项;泰山学者特聘专家(ts201712080);山东省自然科学基金(ZR2017BC082);山东省农业良种工程(2017LZN033;2017LZGC003);山东省现代农业产业技术体系花生遗传育种岗位(SDAIT-04-02);山东省农业科学院农业科技创新工程(CXGC2016A01)

摘　　要：为获得适用于高通量测序的高质量花生叶片DNA,本方法取第三对真叶为实验材料,设计烧制圆球状枪头和离心管装置代替研钵研磨,并在CTAB法的基础上在杂质沉淀前快速分离DNA等简化方法提取DNA。琼脂糖凝胶电泳检测结果表明DNA条带清晰、完整性高。经超微量分光光度计检测,DNA浓度范围在104～131ng/μL,OD_(260)/OD_(280)为1.7～2.0,OD_(260)/OD_(230)大于2.0,说明杂质少,可获得高纯度,高浓度的DNA。此方法与常用的植物基因组DNA提取方法相比,具有成本低、时间短、质量高的优点,可用于基因组重测序、分子生物学技术以及分子标记筛选等后续研究,可提高花生分子育种与筛选工作效率。In order to obtain the high-quality peanut DNA for high-throughput sequencing,the third pair of true leaves were used as experimental material,and the mortar was replaced with ball-shaped pipet and a centrifuge tube.In addition,DNA was isolated from impurities.The concentration of DNA that detected by Nanodrop was 104~131 ng/μl,OD 260/OD 280 was 1.7~2.0,and OD 260/OD 230 was more than 2.0,which indicated that there were few impurities contaminated.Compared with the generally accepted plant genomic DNA extraction method,this method had the advantages of low cost,short time and high quality.It can be applied to the subsequent research such as genome re-sequencing,molecular marker screening,and enhance the efficiency of peanut molecular breeding and screening.

关 键 词：花生 DNA提取 简化方法 

分 类 号：Q781[生物学—分子生物学] S565.203.53[农业科学—作物学]