| 棉花线粒体和线粒体DNA的分离与纯化 |
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| 引用本文： | 刘少林,靖深蓉,严根土,邢朝柱,袁有禄,郭立平,王海林,李建生,郑用连.棉花线粒体和线粒体DNA的分离与纯化[J].棉花学报,1996(6). |
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| 作者姓名： | 刘少林 靖深蓉 严根土 邢朝柱 袁有禄 郭立平 王海林 李建生 郑用连 |
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| 作者单位： | 中国农科院棉花研究所，华中农业大学 |
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| 摘 要： | 本文详细报道了一个针对棉花特点的线粒体以及线粒体ＤＮＡ的抽提方法，即先用蔗糖Ｓｔｅｐ递度和ＤＮａｓｅＩ体外消化分离纯化线粒体，然后氯仿抽提纯化ＤＮＡ。实验过程中采用ＰＶＰ排除棉酚和丹宁的干扰，用ＤＩＥＣＡ抑制酚氧化酶活性，并对许多实验细节作了较大改进。得到的ＤＮＡ可满足酶切、ＰＣＲ等分子生物学分析。
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| 关 键 词： | 棉花 线粒体 线粒体DNA（mtDNA） |
Purification of Cotton Mitochondria and Mitochondrial DNA |
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| Liu Shaolin,Jing Shenrong,Yan Gentu,Xing Chaozhu,Yuan Youlu,Guo Liping,Wang Hailin,Cotton Research Institute CAAS Anyang Li Jiansheng,Zheng Yonglian,Hua Zhoiig,Agricultural University,Wuhan.Purification of Cotton Mitochondria and Mitochondrial DNA[J].Cotton Science,1996(6). |
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| Authors: | Liu Shaolin Jing Shenrong Yan Gentu Xing Chaozhu Yuan Youlu Guo Liping Wang Hailin Cotton Research Institute CAAS Anyang Li Jiansheng Zheng Yonglian Hua Zhoiig Agricultural University Wuhan |
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| Institution: | Liu Shaolin,Jing Shenrong,Yan Gentu,Xing Chaozhu,Yuan Youlu,Guo Liping,Wang Hailin,Cotton Research Institute CAAS Anyang 455112Li Jiansheng,Zheng Yonglian,Hua Zhoiig,Agricultural University,Wuhan 430070 |
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| Abstract: | A method suitable for extraction of cotton mitochondria and mitochondrial DNA(mtDNA)is reported in details.Mitochondria is separated by centrifugation in a sucrosestep density gradient and purified through DNase I digestion. mtDNA is extracted withchloroform. PVP and DIECA are used to bind phenolic compounds and danin and to inacti-vate phenol oxidases,respectively. Many operation steps are greatly improved in this pro-tocol.mtDNA prepared by this method for restriction analvsis,PCR and other molecularoperations were successfully used. |
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| Keywords: | Cotton mitochondria mitochondrial DNA(mtDNA) |
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