| CTAB法提取野野村菌基因组DNA |
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| 引用本文： | 王凡,洪葵.CTAB法提取野野村菌基因组DNA[J].微生物学通报,2010,37(8):1211-1215. |
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| 作者姓名： | 王凡 洪葵 |
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| 作者单位： | 1. 华中农业大学生命科学技术学院,湖北,武汉,430070
2. 华中农业大学生命科学技术学院,湖北,武汉,430070;中国热带农业科学院热带作物生物技术研究所,农业部热带作物生物技术重点开放实验室,海南,海口,571101 |
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| 基金项目： | 国家863计划项目(No. 2007AA09Z415); 国家自然科学基金广东联合基金重点项目(No. U0633008) |
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| 摘 要： | 针对用常规方法难以提取野野村菌基因组DNA的问题,通过选用添加甘氨酸的不同培养基和不同培养时间获得的菌丝体,采用液氮研磨结合CTAB法提取野野村菌DNA,电泳检测及计算OD260/OD280值。结果表明,在添加0.3%甘氨酸的麦芽汁-酵母膏(YE)培养基中振荡培养培养3d的菌丝体适合于DNA提取,用CTAB法获得的基因组DNA,长度约为20kb,且OD260/OD280在1.8左右,达到基因组DNA-DNA杂交的要求。
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| 关 键 词： | 野野村菌属 DNA提取 CTAB法 |
CTAB Method for Genomic DNA Extraction from Nonomuraea |
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| WANG Fan and HONG Kui.CTAB Method for Genomic DNA Extraction from Nonomuraea[J].Microbiology,2010,37(8):1211-1215. |
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| Authors: | WANG Fan and HONG Kui |
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| Institution: | 1. Huazhong Agricultural University of Life Science and Technology, Wuhan, Hubei 430070, China;1. Huazhong Agricultural University of Life Science and Technology, Wuhan, Hubei 430070, China; 2. Key Laboratory of Tropical Crop Biotechnology, Ministry of Agriculture, Institute of Tropical Bioscience and Biotechnology, Chinese Academy of Tropical Agri |
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| Abstract: | In order to obtain genomic DNA which meets the requirements of DNA-DNA hybridization, CTAB method combined with liquid nitrogen grinding was investigated. Cultures from different media added with glycine were harvested in different time. Quality of extracted genomic DNA was assessed by electrophoresis and OD260/OD280 value. Mycelia of shaking culture in yeast extract & malt extract (YE) medium containing 0.3% glycine for 3 days were subjected for DNA extraction. The genomic DNA obtained was about 20 kb, with the OD260/OD280 value around 1.8. |
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| Keywords: | Nonomuraea Genomic DNA extraction CTAB method |
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