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

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。

Abstract: 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 16 S r RNA 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.