传统发酵牦牛酸乳宏基因组DNA三种提取方法比较研究  

Study on comparison of three extraction methods of traditional fermented yak yogurt macro genomic DNA

作　　者：谢婕[1,2] 赵欣[3] 骞宇[3] 陈炼红[4] 李键[4] 索化夷[1,2] 

机构地区：[1]西南大学食品科学学院,重庆400715 [2]西南大学国家食品科学与工程实验教学中心,重庆400715 [3]重庆第二师范学院生物与化学工程系,重庆400067 [4]西南民族大学青藏高原研究院,四川成都610041

出　　处：《食品工业科技》2015年第6期175-178,共4页Science and Technology of Food Industry

基　　金：国家公益性行业(农业)科研专项(201203009);中央高校基本业务费项目(XDJK2013B010);重庆市基础与前沿研究计划项目(CSTC2013JCYJA80006)

摘　　要：微生物区系复杂,自然环境中约95%是无法用常规的培养方法获得的,为了对未能培养或不可培养的微生物进行研究,本实验选用传统发酵牦牛酸乳,分别采用CTAB-SDS-冻融法、液氮研磨法、玻璃珠吸附法来提取宏基因组DNA,然后测得三种方法所提取的DNA的浓度、纯度和片段分布状况,并对总DNA进行16S r RNA基因片段PCR扩增。实验结果表明,本实验所采取的三种DNA提取方法均能提取到适于后续研究质量的DNA,但其中液氮研磨法同时具有提取的总DNA质量较好和步骤简单、成本低的优点,是提取牦牛酸乳宏基因组DNA最合适的方法。Microflora was so complex that around which 95% could not be obtained by culture in conventional way. This paper selected the traditional fermented yak yogurt,and extracted the macro genomic DNA by three methods respectively,including CTAB-SDS-frozen-thawed,liquid nitrogen trituration and glass bead adsorptive process. Then the DNA concentration,purity and fragment distribution,and the total DNA of 16S rRNA gene PCR amplification was tested.The result showed that all the three methods could extract the DNA suitable for the follow-up study. But liquid nitrogen trituration method not only had the priority of a better DNA quality,but also was simplest and lowest costed way compared to the other two methods. So it was the most suitable method to extract yak yogurt macro genomic DNA.

关 键 词：微生物 传统发酵牦牛酸乳 宏基因组 DNA 

分 类 号：TS252[轻工技术与工程—农产品加工及贮藏工程]