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.
An Improved Method for High Quality Metagenomics DNA Extraction from Human and Environmental Samples.
一种从人类和环境样本中提取高质量宏基因组DNA的改进方法
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作者:Bag Satyabrata, Saha Bipasa, Mehta Ojasvi, Anbumani D, Kumar Naveen, Dayal Mayanka, Pant Archana, Kumar Pawan, Saxena Shruti, Allin Kristine H, Hansen Torben, Arumugam Manimozhiyan, Vestergaard Henrik, Pedersen Oluf, Pereira Verima, Abraham Philip, Tripathi Reva, Wadhwa Nitya, Bhatnagar Shinjini, Prakash Visvanathan Gnana, Radha Venkatesan, Anjana R M, Mohan V, Takeda Kiyoshi, Kurakawa Takashi, Nair G Balakrish, Das Bhabatosh
| 期刊: | Scientific Reports | 影响因子: | 3.900 |
| 时间: | 2016 | 起止号: | 2016 May 31; 6:26775 |
| doi: | 10.1038/srep26775 | 种属: | Human |
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