An improved DNA isolation method for metagenomic analysis of the microbial flora of the human intestine
An improved DNA isolation method for metagenomic analysis of the microbial flora of the human intestine
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DOI：
10.1264/jsme2.22.214
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发表时间：
2007-01-01
影响因子：
2.2
通讯作者：
Takami, Hideto
中科院分区：
文献类型：
--
作者：
Morita, Hidetoshi;Kuwahara, Tomomi;Takami, Hideto
使用一种用于宏基因组分析的常规方法以及一种改进方法（该改进方法涉及更高水平的溶菌酶和蛋白酶K，以及添加无色肽酶），对5种严格厌氧和6种兼性厌氧细菌（均为众所周知的人类结肠共生菌）的裂解效率进行了测试。常规方法对10种细菌的裂解效率>80%，而对于所有粪便样本，多枝梭菌JCM 1298(T)的裂解效率为90%。因此，由于通过改进方法分离的DNA样本能够反映微生物群落中近乎真实的基因组信息，我们的改进方法不仅应适用于人类肠道细菌的宏基因组分析，也应适用于其他环境中的细菌。
The efficiency with which lysis of five strictly anaerobic and six facultatively anaerobic bacterial species, all well-known human colonic commensals, were lysed was tested using a reference method for general metagenomic analysis and an improved method that involves higher levels of lysozyme and proteinase K, as well as the addition of achromopeptidase. Ten species were lysed with an efficiency of >80% by the reference method, while the lytic efficiency for Clostridium ramosum JCM 1298(T) was 90%) for all the fecal samples. Accordingly, since the DNA samples isolated by the improved method can reflect nearly true genomic information in the microbial flora, our improved method should be applicable to metagenomic analyses, not only for bacteria in the human intestine but also for bacteria in other environments.