| 引用本文: | 李昂,崔迪,王继华,张斯,杨基先,马放.适用于第2代测序技术的宏基因组DNA提取方法[J].哈尔滨工业大学学报,2012,44(6):20.DOI:10.11918/j.issn.0367-6234.2012.06.005 |
| LI Ang,CUI Di,WANG Ji-hua,ZHANG Si,YANG Ji-xian,MA Fang.Extraction method of metagenomic DNA used for the next generation sequencing technology[J].Journal of Harbin Institute of Technology,2012,44(6):20.DOI:10.11918/j.issn.0367-6234.2012.06.005 |
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| 适用于第2代测序技术的宏基因组DNA提取方法 |
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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
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1.哈尔滨工业大学 城市水资源与水环境国家重点实验室, 150090;2.哈尔滨工业大学 市政环境工程学院, 150090;3.哈尔滨师范大学 生命科学与技术学院, 150025
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| 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169~172．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183,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908％． |
| 关键词: 宏基因组测序 宏基因组DNA 活性污泥 Solexa测序 |
| DOI：10.11918/j.issn.0367-6234.2012.06.005 |
| 分类号:X7031 |
| 基金项目: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) |
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| Extraction method of metagenomic DNA used for the next generation sequencing technology |
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LI Ang1,2, CUI Di1,2, WANG Ji-hua3, ZHANG Si1,2, YANG Ji-xian1,2, MA Fang1,2
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1.State Key Laboratory of Urban Water Resource and Environment, Harbin Institute of Technology, 150090 Harbin, China;2.School of Municipal and Environmental Engineering, Harbin Institute of Technology, 150090 Harbin, China;3.School of Life Science and Technology, Harbin Normal University, 150025 Harbin, China
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| Abstract: |
| 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 169-172. During the sample preparation for the next generation sequencing, D（260／280 nm） of the metagenomic DNA reached to 183 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 908％, which could be used for assemble. |
| Key words: metagenomic sequencing metagenomic DNA the activated sludge Solexa sequencing |
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