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


Ontology type: schema:ScholarlyArticle      Open Access: True


Article Info

DATE

2016-05-31

AUTHORS

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 Prakash, Venkatesan Radha, R. M. Anjana, V. Mohan, Kiyoshi Takeda, Takashi Kurakawa, G. Balakrish Nair, Bhabatosh Das

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. More... »

PAGES

26775

Identifiers

URI

http://scigraph.springernature.com/pub.10.1038/srep26775

DOI

http://dx.doi.org/10.1038/srep26775

DIMENSIONS

https://app.dimensions.ai/details/publication/pub.1033975337

PUBMED

https://www.ncbi.nlm.nih.gov/pubmed/27240745


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