| 芒AFLP反应体系的建立 |
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| 引用本文： | 胡小虎,刁英,郑兴飞,胡晖,余作平,胡中立.芒AFLP反应体系的建立[J].氨基酸和生物资源,2010,32(3):10-14. |
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| 作者姓名： | 胡小虎 刁英 郑兴飞 胡晖 余作平 胡中立 |
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| 作者单位： | 武汉大学生命科学学院,武汉,430072;武汉大学生命科学学院,武汉430072;重庆文理学院生命科学与技术学院,重庆,402168;湖北光芒能源植物有限公司,武汉,430071 |
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| 摘 要： | 以芒DNA为材料,对AFLP分子标记分析中的基因组酶切体系选择性扩增中Mg2+、dNTP和Taq酶浓度等4个因素进行了比较。结果表明20μL基因组双酶切体系中,使用1UEcoRⅠ和1UM seⅠ酶切3 h能够实现完全酶切;选择性扩增的PCR 10μL反应体系中1.4 mmol.L-1Mg2+,0.4 mmol.L-1dNTP及0.6 U Taq酶是进行芒AFLP分析的最佳反应条件,能够得到丰富稳定的带纹。该体系的构建为AFLP技术在芒相关研究中的应用奠定了基础。
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| 关 键 词： | 芒 AFLP 反应体系 |
Establishment of AFLP Analysis System for Miscanthus sinensis |
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| Hu Xiaohu,Diao Ying,Zheng Xingfei,Hu Hui,Yu Zuoping,Hu Zhongli.Establishment of AFLP Analysis System for Miscanthus sinensis[J].Amino Acids & Biotic Resources,2010,32(3):10-14. |
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| Authors: | Hu Xiaohu Diao Ying Zheng Xingfei Hu Hui Yu Zuoping Hu Zhongli |
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| Institution: | Hu Xiaohu1,Diao Ying1,2,Zheng Xingfei1,Hu Hui1,Yu Zuoping3,Hu Zhongli1 (1 College of Life Sciences,Wuhan University,Wuhan 430072,2 College of Life Science , Technology,Chongqing University of Arts , Sciences,Chongqing 402168,3 Hubei GEP Co.Ltd.,Wuhan 430071,China) |
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| Abstract: | By using genomic DNA of Miscanthus sinensis,the concentration of enzymes in DNA double enzyme digestion reaction,and the concentration of Mg2+,dNTP and Taq DNA polymerase in the selective amplification reaction system were optimized in the AFLP analysis system.The results showed that the genomic DNA could be completely digested in 20μL reaction solution containing 1U EcoRⅠand 1U MseⅠin 3hrs,and in 10 μL selective amplification reaction system 1.4mmol·L-1 Mg2+,0.4mmol·L-1 dNTP and 0.6U Taq DNA polymerase were the best for AFLP analysis of M.sinensis.The establishment of the system might lay a foundation for the application of AFLP techniques to the relative research of M.sinensis. |
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| Keywords: | AFLP |
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