| 水稻纹枯菌总DNA提取方法的比较 |
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| 引用本文： | 任衍春,张震,毛雪琴,邱海萍,姜华,柴荣耀,王艳丽,孙国昌.水稻纹枯菌总DNA提取方法的比较[J].浙江农业学报,2011,23(4). |
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| 作者姓名： | 任衍春 张震 毛雪琴 邱海萍 姜华 柴荣耀 王艳丽 孙国昌 |
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| 作者单位： | 1. 浙江师范大学化学与生命科学学院,浙江金华321000;浙江省植物有害生物防控重点实验室一省部共建国家重点实验室培育基地,浙江杭州310021;浙江省农业科学院植物保护与微生物研究所,浙江杭州310021
2. 浙江省植物有害生物防控重点实验室一省部共建国家重点实验室培育基地,浙江杭州310021;浙江省农业科学院植物保护与微生物研究所,浙江杭州310021 |
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| 基金项目： | 国家自然科学基金，浙江省农业科学院科研创新提升项目 |
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| 摘 要： | DNA的浓度,尤其是DNA的纯度严重影响后续基因组酶切等结果.为寻找一种高质量的水稻纹枯菌总DNA的提取方法,采用CTAB法、CTAB改良法、SDS法、蜗牛酶法、蛋白酶K法5种方法分别提取水稻纹枯菌总DNA.通过紫外吸光度测定DNA的纯度和浓度、琼脂糖凝胶电泳、PCR扩增、限制性内切酶酶切反应及Southem杂交评价DNA的质量,最终得出蛋白酶K法是获取高质量水稻纹枯菌基因组DNA的最佳方法.
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| 关 键 词： | 水稻纹枯菌 总DNA Southern杂交 蛋白酶K法 |
Comparison of total DNA extraction methods from Rhizoctonia solani |
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| REN Yan-chun,ZHANG Zhen,MAO Xue-qin,QIU Hai-ping,JIANG Hua,CHAI Rong-yao,WANG Yan-li,SUN Guo-chang.Comparison of total DNA extraction methods from Rhizoctonia solani[J].Acta Agriculturae Zhejiangensis,2011,23(4). |
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| Authors: | REN Yan-chun ZHANG Zhen MAO Xue-qin QIU Hai-ping JIANG Hua CHAI Rong-yao WANG Yan-li SUN Guo-chang |
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| Institution: | REN Yan-chun1,2,3,ZHANG Zhen2,MAO Xue-qin2,QIU Hai-ping2,JIANG Hua2,CHAI Rong-yao2,WANG Yan-li2,*,SUN Guo-chang2,(1College of Chemistry and Life Sciences,Zhejiang Normal University,Jinhua 321000,China,2State Key Laboratory Breeding Base for Zhejiang Sustainable Pest and Disease Control,Hangzhou 310021,3Institute of Plant Protection and Microbiology,Zhejiang Academy of Agricultural Sciences,China) |
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| Abstract: | The DNA quantity and purity strongly limited the digestion efficiency of restrict endonuclease.In order to find an effective method of extracting high quality total DNA from R.solani,the methods,such as CTAB-extraction,SDS-extraction,improved CTAB-extraction,glusulase-extraction and protease K-extraction were compared.The purity and yield of the DNA were determined by ultraviolet absorption spectrum.The DNA quality was also evaluated with agarose gel electrophoresis(AGE),PCR amplification,restrict endonucle... |
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| Keywords: | Rhizoctonia solani total DNA Southern blot protease K-extraction |
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