东北红豆杉RAPD反应体系的优化  

Optimization of RAPD Reaction System of Taxus cuspidata Sieb.et Zucc

作　　者：肖璐[1] 许明子[1] 崔苗[1] 刘博[1] 

机构地区：[1]延边大学农学院农学系,吉林龙井133400

出　　处：《安徽农业科学》2010年第16期8325-8326,8329,共3页Journal of Anhui Agricultural Sciences

摘　　要：[目的]研究东北红豆杉RAPD反应体系的优化条件。[方法]以东北红豆杉叶片为材料,用改进CTAB法提取DNA,分别从模板DNA浓度、引物浓度、dNTP浓度,Taq DNA聚合酶量及镁离子浓度5个方面对东北红豆杉RAPD扩增效果的影响进行分析。[结果]通过各因子的比较分析,建立了东北红豆杉RAPD优化体系:20μl PCR反应总体积中含模板DNA 10 ng,Mg2+2.0 mmol/L,引物15pmolμ/l,dNTPs 0.2 mmol/L,Taq DNA聚合酶1.0 U,10×buffer缓冲液2μl,以双蒸水补充至20μl。[结论]为进一步探索与东北红豆杉性别鉴定相关的研究提供技术支持。[Objective] The research aimed to study the optimization conditions of RAPD reaction system of Taxus cuspidata.[Method] Using the leaves of Taxus cuspidata as materials,DNA was extracted by using modified CTAB method.The influences of template DNA concentration,primer concentration,dNTP concentration,Taq DNA polymerase amount and magnesium ion concentration on the amplification effects of RAPD of Taxus cuspidate were analyzed.[Result] Through analysis and comparison of various factors,the optimized reaction system was established as follows：in a total volume of 20 μl,containing 10 ng DNA,2.0 mmol/L Mg^2＋,0.2 mmol/L dNTPs,15 pmol/μl random primer,1.0 U Taq polymerase,2 μl 10×buffer,and the supplementary ddH2O.[Conclusion] The research provided technical support for further discussion on the ralted studies with the sexual identification of Taxus cuspidate.

关 键 词：东北红豆杉 RAPD 反应条件 

分 类 号：S791.49[农业科学—林木遗传育种]