| 犊牛盲肠微生物总DNA的提取方法 |
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| 引用本文： | 于萍,王加启,卜登攀,邓露芳,刘开朗,魏宏阳,周凌云.犊牛盲肠微生物总DNA的提取方法[J].乳业科学与技术,2009,32(1). |
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| 作者姓名： | 于萍 王加启 卜登攀 邓露芳 刘开朗 魏宏阳 周凌云 |
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| 作者单位： | 中国农业科学院北京畜牧兽医研究所动物营养学国家重点实验室,北京,100193 |
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| 基金项目： | 国际科技合作重点项目，国家支撑计划 |
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| 摘 要： | 采用反复冻融法、玻璃珠法和超声波+反复珠磨的方法提取犊牛盲肠的微生物总DNA,并对以上三种方法进行比较,从中确定最佳的方法,以实现普通实验条件下成功提取符合PCR扩增要求的DNA.经紫外分光光度分析表明,超声波+反复珠磨的方法所得的DNA的A<,260>/A<,280>的比值为1.81,0.8%琼脂糖凝胶电泳结果显示,所提DNA片段分子量大于20 kb,适于酶解和PCR扩增要求.以提取的DNA样品为模板,利用细菌通用引物,对其16S rDNA进行PCR扩增,获得了1.7 kb大小特异性很好的预期条带.这是研究犊牛盲肠微生物的关键一步.
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| 关 键 词： | 犊牛盲肠微生物 细菌总DNA PCR扩增 超声波细胞破碎 反复珠磨法 |
Extraction of Calves Cecum Microbial Total DNA |
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| Yu Ping,Wang Jiaqi,Bu Dengpan,Deng Lufang,Liu Kailang,Wei Hongyang,Zhou Lingyun.Extraction of Calves Cecum Microbial Total DNA[J].JOURNAL OF DAIRY SCIENCE AND TECHNOLOGY,2009,32(1). |
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| Authors: | Yu Ping Wang Jiaqi Bu Dengpan Deng Lufang Liu Kailang Wei Hongyang Zhou Lingyun |
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| Institution: | State Key Laboratory of Animal Nutrition;Institute of Animal Science;Chinese Academy of Agriculture Science;Beijing 100193;P.R.China |
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| Abstract: | In order to extract DNA from calves cecum,three methods(repeated freeze-thaw,bead beat- ing,ultrasionic cell-break method combined with repeated bead beating) were used and compared,re- sults showed that the ultrasionic cell-break method combined with repeated bead beating was the most efficient one.The high-quality total DNA from a wide variety of bacterial specie was isolated and de- termined by a spectrophotometer (A_(260)/A_(280)=1.81) and 0.8% agarose gel electrophoresis.The purified DNA with the molec... |
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| Keywords: | calves cecum microbiota total DNA extraction PCR amplification ultrasionic cell-break repeat bead beating |
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