An Improved Method for High Quality Metagenomics DNA Extraction from Human and Environmental Samples.
An Improved Method for High Quality Metagenomics DNA Extraction from Human and Environmental Samples.
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DOI：
10.1038/srep26775
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发表时间：
2016-05-31
影响因子：
4.6
通讯作者：
Das B
中科院分区：
文献类型：
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作者：
Bag S;Saha B;Mehta O;Anbumani D;Kumar N;Dayal M;Pant A;Kumar P;Saxena S;Allin KH;Hansen T;Arumugam M;Vestergaard H;Pedersen O;Pereira V;Abraham P;Tripathi R;Wadhwa N;Bhatnagar S;Prakash VG;Radha V;Anjana RM;Mohan V;Takeda K;Kurakawa T;Nair GB;Das B
为了通过包括下一代测序在内的高分辨率分子方法探索任何生态系统的天然微生物群落，开发一种灵敏且可重复的DNA提取方法是极其重要的，该方法有助于从生活在该环境中的可培养和未培养的微生物物种中分离足够纯度和数量的微生物DNA。在不损害其基因组的情况下正确裂解异质群落微生物细胞是一个重大挑战。在这项研究中，我们已经开发了一种改进的方法提取社区DNA从不同的环境和人类起源的样品。我们介绍了物理、化学和机械裂解方法的组合，用于适当裂解微生物居民。用盐和有机溶剂沉淀群落微生物DNA。与现有方法进行了比较，证实了本方法的优越性。在不存在大量杂质的情况下，基因组DNA的最大回收率使得该方法便于核酸提取。使用该方法获得的核酸适用于不同的下游应用。这种改进的方法被命名为THSTI方法，以描述开发该方法的研究所。
To explore the natural microbial community of any ecosystems by high-resolution molecular approaches including next generation sequencing, it is extremely important to develop a sensitive and reproducible DNA extraction method that facilitate isolation of microbial DNA of sufficient purity and quantity from culturable and uncultured microbial species living in that environment. Proper lysis of heterogeneous community microbial cells without damaging their genomes is a major challenge. In this study, we have developed an improved method for extraction of community DNA from different environmental and human origin samples. We introduced a combination of physical, chemical and mechanical lysis methods for proper lysis of microbial inhabitants. The community microbial DNA was precipitated by using salt and organic solvent. Both the quality and quantity of isolated DNA was compared with the existing methodologies and the supremacy of our method was confirmed. Maximum recovery of genomic DNA in the absence of substantial amount of impurities made the method convenient for nucleic acid extraction. The nucleic acids obtained using this method are suitable for different downstream applications. This improved method has been named as the THSTI method to depict the Institute where the method was developed.