16S核糖体DNA宏基因组测序中细菌核酸提取方法的比较研究  

Comparative Study of Bacterial Nucleic Extraction Method on 16S r DNA Metagenomic Sequencing

作　　者：王伟[1,2] 王玉琢[3] 舒鹏[1,2] 米志强[2] 安小平[2] 裴广倩[2] 刘文丽[2] 袁文俊[3] 史套兴[1,2] 童贻刚[1,2] 

机构地区：[1]安徽医科大学,安徽合肥230032 [2]军事医学科学院微生物流行病研究所,北京100071 [3]宁夏医科大学,宁夏银川750000

出　　处：《生物技术通讯》2015年第4期551-555,共5页Letters in Biotechnology

基　　金：国家科技重大专项(2013ZX10004605;2011ZX10004001;2013ZX10004607-004;2013ZX10004217-002-003);国家高技术研究发展计划(2012AA022003;2014AA021402)

摘　　要：目的：探究细菌核酸提取方法对16S核糖体DNA（r DNA）宏基因组测序的影响。方法：分别采用TRIzol提取、试剂盒提取、溶葡酶＋试剂盒提取、超声波＋试剂盒提取、超声波＋溶葡酶＋试剂盒提取、匀浆＋试剂盒提取、匀浆＋溶葡酶＋试剂盒提取共7种方法对模拟样本进行细菌核酸提取,特异性地扩增16S r DNA的V1~V2高变区,测序后分析模拟样本中各种细菌的含量和相对分布。结果：超声波处理结合试剂盒提取方法中不同细菌数据分布及含量都相对均匀,是一种广谱、高效的细菌核酸提取方法。结论：超声波处理结合试剂盒提取方法适用于细菌宏基因组测序中样本核酸的提取。Objective：To exploring bacterial nucleic extraction method on 16 S ribosomal DNA（r DNA） metagenom-ic sequencing.Methods：Bacterial nucleic acids were extracted separately by TRIzol,DNA extraction kit,lysozyme＋ DNA extraction kit,ultrasonic ＋ DNA extraction kit,ultrasonic ＋ lysozyme ＋ DNA extraction kit,homogenate ＋DNA extraction kit and homogenate ＋ lysozyme ＋ DNA extraction kit,totally seven different kinds of methods.Af-ter sequencing,16 S r DNA V1~V2 hypervariable region was amplified specifically.After sequencing,the content ofvarious bacteria and relative distribution of simulation samples were analyzed.Results：The results showed that da-ta distribution and contents are relatively uniform in ultrasonic extraction method combined with kit different,itturns out a broad-spectrum,highly efficient bacterial DNA extracting method.Conclusion：Ultrasonic extractionmethod is suitable for extracting the bacteria nucleic acid in 16 S r DNA metagenomic sequencing.

关 键 词：16S核糖体DNA 核酸提取 基因组测序 

分 类 号：Q78[生物学—分子生物学]