西蓝薹RAPD-PCR反应体系的建立与优化  

Establishment and optimization of RAPD-PCR reaction system in Broccolini

作　　者：尹彩霞[1,2] 张继栋[1,2] 李向阳 刘自珠 张华 张金艳[1,2] 乔爱民[1,2] 

机构地区：[1]仲恺农业工程学院园艺园林学院,广东广州510225 [2]广东省教育厅热带亚热带花卉与园林植物重点实验室,广东广州510225 [3]广州市农业科学研究院,广东广州510308

出　　处：《仲恺农业工程学院学报》2010年第3期22-26,共5页Journal of Zhongkai University of Agriculture and Engineering

基　　金：广东省科技计划项目(2007A020200004-9);广州市蔬菜育种新技术应用研究重点实验室开放基金(D1081229)资助项目

摘　　要：以新型蔬菜西蓝薹为研究对象,采用改良的十六烷基三甲基溴化铵(CTAB)法提取基因组DNA,并通过单因素多水平梯度试验,筛选了Taq酶、引物、DNA模板、Mg2+、dNTPs的浓度或用量,建立了西蓝薹RAPD-PCR的最佳反应体系:25μL反应体系中含0.8 UTaq酶、0.40μmol/L引物、30 ng DNA模板、1.6 mmol/LMg2+、0.25 mmol/L dNTPs、2.5μL 10×PCR buffer.利用优化的反应体系,对5个西蓝薹品种进行体系稳定性检测,结果表明该反应体系的重复性和稳定性良好.The genomic DNA of new vegetable broccolini was extracted by modified CTAB method.The factors affecting RAPD results were studied through the ladder experiments of single factor and multilevel,including concentrations or dosages of Taq polymerase,primer,template DNA,Mg2＋and dNTPs.An optimal reaction system（25 μL）was established which contained 0.8U Taq polymerase,0.40 μmol/L primer,30 ng template DNA,1.6 mmol/L Mg2＋,0.25 mmol/L dNTPs,and 2.5 μL 10×PCR buffer.Five broccolini cultivars were used to test the stabilization of the optimized reaction system.The results showed that the optimized reaction system of RAPD-PCR for the new vegetable was stable and repeatable.

关 键 词：西蓝薹 RAPD 反应体系 优化 

分 类 号：S634.9[农业科学—蔬菜学]