| 柳叶蜡梅RAPD反应体系的研究 |
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| 引用本文： | 杨华,张都海,陈磊,杜国坚.柳叶蜡梅RAPD反应体系的研究[J].江西林业科技,2010(5):18-20. |
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| 作者姓名： | 杨华 张都海 陈磊 杜国坚 |
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| 作者单位： | [1]浙江省林业科学研究院,浙江杭州310023 [2]浙江科技学院,浙江杭州310023 |
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| 基金项目： | 浙江省属科研院所专项公益技术攻关项目 |
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| 摘 要： | 以柳叶蜡梅（Chimonanthus Salicfiolius Hu）为材料,对其DNA提取方法和PCR扩增反应体系的建立和优化进行探讨。最适宜的PCR反应条件为：20μL PCR反应体积中含有1×缓冲液,20 ng模板DNA,2.75 mmol/LMgCl2,0.25 mmol/L dNTPs,2.0 U Taq DNA聚合酶,0.4μmol/L引物。扩增程序为：94℃预变性2 min;再32个循环：94℃60 s,36℃30 s和72℃120 s;最后72℃延伸10 min。
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| 关 键 词： | 柳叶蜡梅 RAPD 优化 |
Optimization of RAPD Reaction System for Chimonanthus salicfiolius |
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| YANG Hua,ZHANG Duhai,CHEN Lei,DU Guojian.Optimization of RAPD Reaction System for Chimonanthus salicfiolius[J].Jiangxi Forestry Science and Technology,2010(5):18-20. |
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| Authors: | YANG Hua ZHANG Duhai CHEN Lei DU Guojian |
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| Institution: | 1.Zhejiang Academy of Forestry,Hangzhou Zhejiang 310023,China;2.Zhejiang University of Science and Technology,Hangzhou Zhejiang 310023,China) |
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| Abstract: | Taking Chimonanthus salicfiolius Hu as molecular study material,the building and optimization of its DNA extraction method and PCR Amplification Reaction System were discussed.The results showed that the conditions being suitable for RAPD-PCR as follows： 1×buffer,20 ng template DNA,2.75 mmol/L MgCl2,0.25 mmol/L dNTPs,2.0 U Taq DNA polymerase,0.4 μmol/L primer.The amplification cycling was 1 cycle at 94 ℃ for 2 min;32 cycles at 94 ℃ for 60 s,36 ℃ for 30 s,and 72 ℃ for 120 s,respectively;and 1 cycle at 72 ℃ for 10 min. |
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| Keywords: | Chimonanthus salicfiolius Hu RAPD Optimization |
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