| 药用植物胡黄连ISSR-PCR反应体系的建立与优化 |
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| 引用本文： | 刘小莉,普春霞,杨耀文,钱子刚.药用植物胡黄连ISSR-PCR反应体系的建立与优化[J].时珍国医国药,2010,21(2). |
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| 作者姓名： | 刘小莉 普春霞 杨耀文 钱子刚 |
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| 作者单位： | 1. 云南中医学院·中药学院,云南,昆明,650500;云南大学生命科学学院,云南,昆明,650091
2. 云南中医学院·中药学院,云南,昆明,650500 |
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| 基金项目： | 国家科技基础性工作专项重点项目子课题，云南中医学院科学研究基金项目 |
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| 摘 要： | 目的 优选胡黄连稳定的ISSR-RCR反应体系.方法 采用4因素(dNTRs、Mg~(2+)、Taq酶、引物)4水平的正交实验和单因子梯度优化相结合的方法.结果 适于胡黄连的25μl ISSR-PCR反应体系中各因素最佳浓度分别为10×buffer 2.5 μl,dNTPs 150μmol/L,Mg~(2+)1.5mmol/L,Taq酶1U,引物0.5 μmol/L,DNA 20 ng或10×buffer 2.5 μl,dNTPs 150μmol/L,Mg~(2+)2 mmol/L,Taq酶0.5 U,引物0.4 μmol/L,DNA 20 ng.结论 对于胡黄连ISSR-RCR试验,一次正交实验完全可以建立起满足要求的PCR体系.
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| 关 键 词： | 胡黄连 正交实验 单因子 |
Establishment and Optimization of ISSR PCR Reaction System for Medicinal Plant Neopicrorhiza scrophulariiflora |
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| LIU Xiao-li,PU Chun-xia,YANG Yao-wen,QIAN Zi-gang.Establishment and Optimization of ISSR PCR Reaction System for Medicinal Plant Neopicrorhiza scrophulariiflora[J].Lishizhen Medicine and Materia Medica Research,2010,21(2). |
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| Authors: | LIU Xiao-li PU Chun-xia YANG Yao-wen QIAN Zi-gang |
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| Institution: | LIU Xiao-li1,2,PU Chun-xia1,YANG Yao-wen1,QIAN Zi-gang1(1.Department of Chinese Materia Medica,Yunnan College of Traditional Chinese Medicine,Kunming,650500,China,2.College of Life Science,Yunnan University,650091 China) |
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| Abstract: | Objective To optimize the stable ISSR reaction system for Neopicrorhiza scrophulariiflora.Methods Concentration of dNTPs,Mg~(2+),Taq DNA polymerase and primers were studied by orthogonal experiment combined with single factor experiment.Results The optimal ISSR-PCR reaction system(25μl) was as follows:10×buffer 2.5 μl,dNTPs150 μmol/L,Mg~(2+)1.5mmol/L,Taq DNA polymerase 1U,primers 0.5 μmol/L,DNA 20 ng or 10×buffer 2.5 μl,dNTPs 150 μmol/L,Mg~(2+) 2mmol/L,Taq DNA polymerase 0.5U,primers 0.4 μmol/L,DNA 20 ng.Conclusion For Neopicrorhiza scrophulariiflora,one orthogonal experiment can establish satisfactory PCR system. |
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| Keywords: | ISSR-PCR Neopicrorhiza scrophulariiflora Orthogonal experiment Single factor experiment ISSR-PCR |
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