| 濒危药用植物延龄草RAPD反应体系的建立 |
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| 引用本文： | 梁薇,何正权,胡勇刚,李雪萍,陈发菊,姚伟,梁宏伟.濒危药用植物延龄草RAPD反应体系的建立[J].福建林业科技,2007,34(1):14-17,100. |
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| 作者姓名： | 梁薇 何正权 胡勇刚 李雪萍 陈发菊 姚伟 梁宏伟 |
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| 作者单位： | 1. 三峡大学生物技术研究中心、天然产物研究与利用湖北省重点实验室,湖北,宜昌,443002
2. 三峡大学生物技术研究中心、天然产物研究与利用湖北省重点实验室,湖北,宜昌,443002;北京林业大学生物科学与技术学院,北京,100083 |
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| 摘 要： | 以濒危药用植物延龄草(Trillium tschonoskiiM.)的叶片为材料,采用改良的CTAB法提取基因组DNA进行RAPD扩增,优化了RAPD扩增反应的程序及模板、引物、dNTP、Taq酶浓度等参数。结果表明:适合延龄草的RAPD扩增程序为:94℃预变性5 min,然后执行40个循环,每个循环中94℃变性1 min,36℃退火1 min,72℃延伸2 min,最后72℃延伸10min;适宜的反应体系为:25μL总体积中含DNA模板50 ng,Mg2+2.5 mM,引物0.6μM,dNTP 0.2 mM,Taq DNA聚合酶3 U。
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| 关 键 词： | 延龄草 RAPD 反应体系 |
| 文章编号： | 1002-7351（2007）01-0014-04 |
| 修稿时间： | 2006-06-092006-10-05 |
Optimization of RAPD reaction system for endangered pharmacy plant Trillium Tschonoskii |
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| LIANG Wei,HE Zheng-quan,HU Yong-gang,LI Xue-ping,CHEN Fa-ju,YAO Wei,LIANG Hong-wei.Optimization of RAPD reaction system for endangered pharmacy plant Trillium Tschonoskii[J].Journal of Fujian Forestry Science and Technology,2007,34(1):14-17,100. |
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| Authors: | LIANG Wei HE Zheng-quan HU Yong-gang LI Xue-ping CHEN Fa-ju YAO Wei LIANG Hong-wei |
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| Institution: | 1. Key Lab of Natural Product Research and Utility, Biotechnology Research Center, China Three Gorges University, Yichang, Hubei 443002, China; 2. School of Biological Science and Biotechnology, Beijing Forestry University, Beijing 100083, China. |
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| Abstract: | The genomic DNA was extracted from young leaves of endangered medicinal plant Trillium Tschonoskii by means of improved CTAB method.The RAPD reaction system for Trillium Tschonoskii was optimized including cycling program and the concentration of template,primer,dNTP,Taq DNA polymerase,and Mg2+.The result indicated that the optimal cycling program was 5 min at 94℃ for predenaturation;then 40 cycles of 1 min at 94℃ for denaturation,1 min at 36℃ for annealing,2 min at 72℃ for extension;finally extension at 72℃ for 8 min.The RAPD reaction system was that template 50ng,Mg2+ 2.5 mM,primer 0.6 μM,dNTP 0.2 mM, Taq DNA polymerase 3U. |
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| Keywords: | Trillium tschonoskii Maxim RAPD reaction system |
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