适用于第2代测序技术的宏基因组DNA提取方法  

Extraction method of metagenomic DNA used for the next generation sequencing technology

作　　者：李昂[1,2] 崔迪[1,2] 王继华[3] 张斯[1,2] 杨基先[1,2] 马放[1,2] 

机构地区：[1]哈尔滨工业大学城市水资源与水环境国家重点实验室,哈尔滨150090 [2]哈尔滨工业大学市政环境工程学院,哈尔滨150090 [3]哈尔滨师范大学生命科学与技术学院,哈尔滨150025

出　　处：《哈尔滨工业大学学报》2012年第6期20-23,共4页Journal of Harbin Institute of Technology

基　　金：国家创新研究群体科学基金资助项目(51121062);国家自然科学基金资助项目(51108120;51178139;51108145);第46批中国博士后科学基金(20090460901);第4批中国博士后科学基金特别资助(201104430);水体污染控制与治理国家重大专项子课题(2012ZX07301001)

摘　　要：为快速、高效、大量地获得城市污水处理厂活性污泥中的宏基因组DNA,将3种不同的基因组DNA提取方式进行对比.结果显示,采用液氮研磨+基因组提取试剂盒相结合的方法获得的宏基因组DNA信息量最大,质量好,D(260/280 nm)在1.69～1.72.在进行高通量测序前的样品制备过程中,样品经过初步处理后宏基因组D(260/280 nm)达1.83,满足文库构建要求.利用Solexa Genome Analyzer System测序结果显示,两个样品正向测序结果良好,可用于序列拼接的高质量数据条数占90.8%.Extraction of metagenomic DNA from environmental sample was the main restrictive factor for the metagenomic sequencing.Three different extraction methods were used and compared in this study.The results indicated that the metagenomic DNA obtained from the activated sludge by Liquid Nitrogen Grinding ＋ DNA Extraction Kit was the best sample due to the largest information and best quality,and D（260 /280 nm） was between 1.69-1.72.During the sample preparation for the next generation sequencing,D（260 /280 nm） of the metagenomic DNA reached to 1.83 after purification,which met the requirement of library construction.After the sequencing by Solexa Genome Analyzer System,the results showed that the forward sequencing of these two samples was successful,and the high quality data was about 90.8%,which could be used for assemble.

关 键 词：宏基因组测序 宏基因组DNA 活性污泥 Solexa测序 

分 类 号：X703.1[环境科学与工程—环境工程]