An Improved Method for High Quality Metagenomics DNA Extraction from Human and Environmental Samples

Satyabrata Bag, Bipasa Saha, Ojasvi Mehta, D Anbumani, Naveen Kumar, Mayanka Dayal, Archana Pant, Pawan Kumar, Shruti Saxena, Kristine H Allin, Torben Hansen, Manimozhiyan Arumugam, Henrik Vestergaard, Oluf Pedersen, Verima Pereira, Philip Abraham, Reva Tripathi, Nitya Wadhwa, Shinjini Bhatnagar, Visvanathan Gnana PrakashVenkatesan Radha, R M Anjana, V Mohan, Kiyoshi Takeda, Takashi Kurakawa, G Balakrish Nair, Bhabatosh Das

131 Citationer (Scopus)

Abstract

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.

OriginalsprogEngelsk
TidsskriftScientific Reports
Vol/bind6
Sider (fra-til)26775
ISSN2045-2322
DOI
StatusUdgivet - 31 maj 2016

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