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

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 quant...

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Autores principales: 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
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Nature Publishing Group 2016
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4886217/
https://www.ncbi.nlm.nih.gov/pubmed/27240745
http://dx.doi.org/10.1038/srep26775
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author 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
author_facet 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
author_sort Bag, Satyabrata
collection PubMed
description 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.
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spelling pubmed-48862172016-06-08 An Improved Method for High Quality Metagenomics DNA Extraction from Human and Environmental Samples 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 Sci Rep Article 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. Nature Publishing Group 2016-05-31 /pmc/articles/PMC4886217/ /pubmed/27240745 http://dx.doi.org/10.1038/srep26775 Text en Copyright © 2016, Macmillan Publishers Limited http://creativecommons.org/licenses/by/4.0/ This work is licensed under a Creative Commons Attribution 4.0 International License. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in the credit line; if the material is not included under the Creative Commons license, users will need to obtain permission from the license holder to reproduce the material. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/
spellingShingle Article
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
An Improved Method for High Quality Metagenomics DNA Extraction from Human and Environmental Samples
title An Improved Method for High Quality Metagenomics DNA Extraction from Human and Environmental Samples
title_full An Improved Method for High Quality Metagenomics DNA Extraction from Human and Environmental Samples
title_fullStr An Improved Method for High Quality Metagenomics DNA Extraction from Human and Environmental Samples
title_full_unstemmed An Improved Method for High Quality Metagenomics DNA Extraction from Human and Environmental Samples
title_short An Improved Method for High Quality Metagenomics DNA Extraction from Human and Environmental Samples
title_sort improved method for high quality metagenomics dna extraction from human and environmental samples
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4886217/
https://www.ncbi.nlm.nih.gov/pubmed/27240745
http://dx.doi.org/10.1038/srep26775
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