| 桉树随机引物PCR反应体系优化研究 |
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| 引用本文： | 邓紫宇,张照远,项东云,唐庆兰,陈健波,卢翠香,叶露.桉树随机引物PCR反应体系优化研究[J].广西林业科技,2012(1):23-26. |
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| 作者姓名： | 邓紫宇 张照远 项东云 唐庆兰 陈健波 卢翠香 叶露 |
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| 作者单位： | [1]广西大学林学院,南宁530000 [2]广西林业科学研究院国家林业局中南速生材繁育实验室广西优良用材林资源培育重点实验室,南宁530002 [3]南京林业大学森林资源与环境学院,南京210037 |
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| 基金项目： | 广西林科院基本科研业务费项目（林科200907号）; 广西科技攻关与新产品试制项目（桂科攻1123004-3A）; 广西林业科技攻关项目（“十五”林科字〔2000〕15号） |
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| 摘 要： | 以桉树DNA为模板,采用单因素试验方法研究dNTP浓度、Mg2＋浓度及退火温度对桉树随机引物PCR反应结果的影响。试验结果表明,最优的反应体系为：20μL的PCR反应体积中,10×Buffer2.0μL,Mg2＋（25 mmol/L）1.28μL,dNTP（10 mmol/L）0.4μL,Primer（10μmol/L）2.0μL,Taq（5 U/μL）0.2μL,DNA2.0μL;最优扩增程序：94℃预变性2 min,然后进行35个循环（94℃变性30 s,36℃退火30 s,72℃延伸2 min）,最后72℃延伸10 min。
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| 关 键 词： | 桉树 随机引物PCR 体系优化 |
The Study on Optimization of Arbitrary-primer PCR Reaction System in Eucalypt |
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| Authors: | DENG Zi-yu ZHANG Zhao-yuan XIANG Dong-yun TANG Qing-lan CHEN Jian-bo LU Cui-xiang YE Lu |
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| Institution: | 1.Forestry College of Guangxi University,Nanning 530000,China;2.Guangxi ForestryResearch Institute,Key Laboratory of Central South Fast-growing Timber Cultivation of Forestry Ministryof China,Guangxi Key Laboratory of Superior Timber Trees Resource Cultivation,Nanning 530002,China;3.College of Forest Resources and Environment of Nanjing Forestry University, Nanjing 210037,China) |
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| Abstract: | Using DNA of Eucalypt as template,the effect of dNTP concentration,Mg2＋ concentration and annealing temperature on the response of Arbitrary-primer PCR was examined by applying single factor experimenting method.The results demonstrated that the optimal condition of reaction for Eucalypt was 2.0 μL Buffer（10 x）,1.28 μL Mg2＋（25 mmol/L）,0.4 μL dNTP（10 mmol/L）,2.0 μL Primer（10 μmol/L）,0.2 μL Taq（5 U/μL）,2.0 μL DNA.The optimization of the amplification process consisted of a pre-denaturation step at 94℃ for 2 minutes,followed by 35 cycles of denaturation at 94℃/30 s,annealing at 36℃/30 s and extension at 72℃/2 min,plus a final extension of 10 min at 72°C. |
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| Keywords: | eucalypt arbitrary-primer PCR reaction system optimization |
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