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

摘要

目的：探究细菌核酸提取方法对16S核糖体DNA（rDNA）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 rDNA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 16S ribosomal DNA(rDNA) 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, 16S rDNA V1~V2 hypervariable region was amplified specifically. After sequencing, the content of various 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, it turns out a broad-spectrum, highly efficient bacterial DNA extracting method. Conclusion: Ultrasonic extraction method is suitable for extracting the bacteria nucleic acid in 16S rDNA metagenomic sequencing.