| 烟草RAPD反应体系的建立与优化研究 |
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| 引用本文： | 杨友才,周清明,尹晗琪.烟草RAPD反应体系的建立与优化研究[J].中国农学通报,2005,21(5):97-97. |
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| 作者姓名： | 杨友才 周清明 尹晗琪 |
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| 作者单位： | 湖南农业大学烟草工程技术研究中心,湖南长沙,410128 |
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| 基金项目： | 湖南省科技厅重大科技攻关项目(01NKY1002)。 |
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| 摘 要： | 以烤烟品种为材料，研究了烟草RAPD分析过程中的影响因素，包括模板浓度、Mg2+、dNTP、引物、Taq 酶、循环次数、退火温度等，建立了适于烟草RAPD分析的PCR反应体系：即在25μl反应体系中，模板用量为40ng；引物浓度为0.4μM；Mg2+浓度为2.5mM；dNTP浓度为0.2mM；Taq DNA聚合酶用量为1U。扩增程序为94℃预变性5min；然后94℃变性1min，38℃复性1min，72℃延伸1.5min，39个循环；最后72℃延伸5min。
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| 关 键 词： | 烟草 RAPD 反应体系 建立与优化 |
Establishment and Optimization of RAPD Reaction System in Tobacco |
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| Yang Youcai,Zhou Qingmin,Yin Hanqi.Establishment and Optimization of RAPD Reaction System in Tobacco[J].Chinese Agricultural Science Bulletin,2005,21(5):97-97. |
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| Authors: | Yang Youcai Zhou Qingmin Yin Hanqi |
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| Institution: | (Tobacco Engineering and Technology Research Center, HNAU, Changsha 410128) |
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| Abstract: | The factors influencing RAPD Analysis, including the concentration of DNA template, Mg2+, dNTP, primers, Taq polymerase, Thermal cycles and annealing temperature in tobacco were studied. An optimal PCR system for RAPD in tobacco was found: in 25μl reaction solution, contained 40ng DNA template, 0.4μM random primers, 2.5mM Mg2+, 0.2mM dNTP, 1U Taq polymerase. The amplification program was devised: 94℃ for 5min;denaturing at 94℃ for 1min; primer annealing at 38℃ for 1min, extension at 72℃ for 1.5min, 39 cycles; at last extension at 72℃ for 5min. |
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| Keywords: | Tobacco RAPD Reaction system Establishment and optimization |
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