摘要
对浙江省天台山自然香果树种群的DNA进行了提取和RAPD扩增条件的优化,分别测试了镁离子、dNTP、模板DNA含量、引物浓度、DNA聚合酶量和牛血清白蛋白浓度对反应结果的影响,通过各因子的组合研究,可知香果树RAPD分析较适宜的扩增条件为:15μLPCR反应体积,1×Taq酶配套缓冲液,1 8mmol·L-1MgCl2,2UTaq酶;10ng模板DNA;20pmol引物;dATP、dCTP、dGTP、dTTP各0 1mmol·L-1。以此适宜条件对12个引物进行扩增,共扩增出51个DNA片段,多态位点百分率为45 1%,种群内遗传多样性较低,进一步研究不同种群香果树的遗传多样性具有一定的意义。
The optimal conditions of RAPD amplification of Emmenopterys henryi from Tiantai mountain in Zhejiang province was screened.After the analysis of the content of Mg^(2+),dNTP,DNA templates,primers and DNA polymerase,the optimal conditions of RAPD was determined as follows:1×Taq polymerase corresponding buffer,1.8 mmol·L^(-1) MgCl_2,2U Taq DNA polymerase,10 ng template DNA,20 pmol primer,0.1 mmol·L^(-1) dATP,dCTP,dGTP,dTTP for each in total 15μL reaction volume.12 random primers were selected in the amplification and 51 repetitive loci were produced.The percentage of polymorphic loci was 45.1% and the genetic diversity within population of E.henryi was relatively low,as indicated by Shannon index of phenotypic diversity and Nei index.It was significance for the further study on the genetic diversity of E.henryi from different populations.
出处
《福建林业科技》
2004年第2期36-40,共5页
Journal of Fujian Forestry Science and Technology
基金
浙江省教育厅科研计划项目(2002005)
