小麦籽粒DNA提取方法的比较及SSR反应体系的优化研究  

Comparison of DNA Extraction Methods from Wheat Grain and the Optimization of SSR System

作　　者：丁海燕[1] 丁晨[1] 

机构地区：[1]大庆师范学院生物工程学院,黑龙江大庆163712

出　　处：《麦类作物学报》2016年第3期287-291,共5页Journal of Triticeae Crops

基　　金：黑龙江省教育厅科学技术研究项目(12513004);大庆师范学院科学研究基金项目(12ZR04);大庆市科技局科技计划项目(szdfy-2015-65)

摘　　要：为了快速获得较高质量的DNA用于SSR分析,并建立一套稳定的SSR反应体系,以小麦单粒干种子为材料,比较了CTAB法和SDS法获得的DNA条带清晰度和RNA残留带的清晰度;同时,研究了模板DNA浓度、引物浓度、dNTPs浓度以及Taq酶用量对SSR结果的影响。结果表明,CTAB法微量提取小麦干种子DNA可获得理想的扩增结果;SSR的最佳反应体系是：模板DNA浓度为4.0～4.8ng·μL^（-1）,Taq酶用量为1U,引物浓度为50～200nmol·L^（-1）,dNTPs浓度为50～500μmol·L^（-1）。In order to obtain high quality DNA for SSR analysis and to establish a stable SSR technology system,DNA extraction and optimal conditions of SSR procedure were studied by using single dry grain of wheat as experimental material.The definition of DNA band and RNA residual band extracted with CTAB and SDS methods were compared.The effects of DNA concentration,primer concentration,dNTPs concentration and Taqenzyme concentration on the results of SSR were investigated.The results showed that the effective amplification products could be obtained with the DNA templates extracted from dry grain with CTAB buffer.SSR optimal reaction system was：4.0-4.8ng·μL^（-1） of template,1Uof Taq emzyme,50-200nmol·L^（-1） of primer,and 50-500μmol·L^（-1） of dNTPs.

关 键 词：小麦 种子 CTAB法 SDS法 SSR 

分 类 号：S512.1[农业科学—作物学] S330