| 水稻RAPD反应体系的正交优化 |
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| 引用本文： | 张安世,邢智峰,徐九文,张利民,韦慧彦.水稻RAPD反应体系的正交优化[J].生物技术通报,2009(12). |
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| 作者姓名： | 张安世 邢智峰 徐九文 张利民 韦慧彦 |
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| 作者单位： | 1. 焦作师范高等专科学校生物系,焦作,454003
2. 焦作农业科学研究院,焦作,454003 |
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| 摘 要： | 以焦旱1号总DNA为材料,首先对影响RAPD-PCR反应的模板DNA、Mg~(2+)、dNTP、引物和Taq DNA聚合酶浓度等因素进行了初步优化,分析了各因素对RAPD-PCR扩增结果的影响.在此基础上对影响RAPD-PCR反应的Mg~(2+)、dNTP、引物和Taq DNA聚合酶浓度等4个主要因素进行正交优化,研究结果表明:在25 μl RAPD-PCR反应体系中,模板DNA 20 ng;Mg~(2+)浓度1.5mmol/L;dNTP的浓度0.2mmol/L;引物量15 pmol;Taq DNA聚合酶1.0 U.在此最佳条件下,利用引物B8对18个北方粳稻品种进行了成功的扩增.
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| 关 键 词： | 水稻 优化 反应体系 |
Orthogonal Optimization of RAPD Reaction System in Rice |
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| Zhang Anshi,Xing Zhifeng,Xu Jiuwen,Zhang Limin,Wei Huiyan.Orthogonal Optimization of RAPD Reaction System in Rice[J].Biotechnology Bulletin,2009(12). |
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| Authors: | Zhang Anshi Xing Zhifeng Xu Jiuwen Zhang Limin Wei Huiyan |
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| Abstract: | It was to obtain the optimum RAPD-PCR reaction system for Jiaohan No.1,one test was designed to screen the suitable concentration of 4 factors(Mg~(2+),dNTP,primer,Taq)in the reaction system.The optimal protocol was accomplished by orthogonal method in 25 μl reaction volumes containing 20 ng template DNA,1.5 mmol/L MgCl_2,0.2 mmol/L dNTPs,1.5 pmol each primer and 1.0 unit Tag polymerase.The results showed that primers B8 can be used to amplify 18 species of Japonica Rice. |
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| Keywords: | RAPD PCR Rice RAPD PCR Optimize Reaction system |
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