title: An Improved Method for High Quality Metagenomics DNA Extraction from Human and Environmental Samples.
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.
authors.affiliation: Molecular Genetics Laboratory, Centre for Human Microbial Ecology, Translational Health Science and Technology Institute, NCR Biotech Science Cluster, Faridabad, India.
authors.forename: Satyabrata
authors.initials: S
authors.lastname: Bag
authors.affiliation: Molecular Genetics Laboratory, Centre for Human Microbial Ecology, Translational Health Science and Technology Institute, NCR Biotech Science Cluster, Faridabad, India.
authors.forename: Bipasa
authors.initials: B
authors.lastname: Saha
authors.affiliation: Molecular Genetics Laboratory, Centre for Human Microbial Ecology, Translational Health Science and Technology Institute, NCR Biotech Science Cluster, Faridabad, India.
authors.forename: Ojasvi
authors.initials: O
authors.lastname: Mehta
authors.affiliation: Molecular Genetics Laboratory, Centre for Human Microbial Ecology, Translational Health Science and Technology Institute, NCR Biotech Science Cluster, Faridabad, India.
authors.forename: D
authors.initials: D
authors.lastname: Anbumani
authors.affiliation: Molecular Genetics Laboratory, Centre for Human Microbial Ecology, Translational Health Science and Technology Institute, NCR Biotech Science Cluster, Faridabad, India.
authors.forename: Naveen
authors.initials: N
authors.lastname: Kumar
authors.affiliation: Molecular Genetics Laboratory, Centre for Human Microbial Ecology, Translational Health Science and Technology Institute, NCR Biotech Science Cluster, Faridabad, India.
authors.forename: Mayanka
authors.initials: M
authors.lastname: Dayal
authors.affiliation: Molecular Genetics Laboratory, Centre for Human Microbial Ecology, Translational Health Science and Technology Institute, NCR Biotech Science Cluster, Faridabad, India.
authors.forename: Archana
authors.initials: A
authors.lastname: Pant
authors.affiliation: Molecular Genetics Laboratory, Centre for Human Microbial Ecology, Translational Health Science and Technology Institute, NCR Biotech Science Cluster, Faridabad, India.
authors.forename: Pawan
authors.initials: P
authors.lastname: Kumar
authors.affiliation: Molecular Genetics Laboratory, Centre for Human Microbial Ecology, Translational Health Science and Technology Institute, NCR Biotech Science Cluster, Faridabad, India.
authors.forename: Shruti
authors.initials: S
authors.lastname: Saxena
authors.affiliation: Section of Metabolic Genetics, Novo Nordisk Foundation Center for Basic Metabolic Research, University of Copenhagen, Copenhagen, Denmark.
authors.forename: Kristine H
authors.initials: KH
authors.lastname: Allin
authors.affiliation: Section of Metabolic Genetics, Novo Nordisk Foundation Center for Basic Metabolic Research, University of Copenhagen, Copenhagen, Denmark.
authors.forename: Torben
authors.initials: T
authors.lastname: Hansen
authors.affiliation: Section of Metabolic Genetics, Novo Nordisk Foundation Center for Basic Metabolic Research, University of Copenhagen, Copenhagen, Denmark.
authors.forename: Manimozhiyan
authors.initials: M
authors.lastname: Arumugam
authors.affiliation: Section of Metabolic Genetics, Novo Nordisk Foundation Center for Basic Metabolic Research, University of Copenhagen, Copenhagen, Denmark.
authors.forename: Henrik
authors.initials: H
authors.lastname: Vestergaard
authors.affiliation: Section of Metabolic Genetics, Novo Nordisk Foundation Center for Basic Metabolic Research, University of Copenhagen, Copenhagen, Denmark.
authors.forename: Oluf
authors.initials: O
authors.lastname: Pedersen
authors.affiliation: Department of Gastroenterology, P.D. Hinduja Hospital and Medical Research Centre, Mumbai, India.
authors.forename: Verima
authors.initials: V
authors.lastname: Pereira
authors.affiliation: Department of Gastroenterology, P.D. Hinduja Hospital and Medical Research Centre, Mumbai, India.
authors.forename: Philip
authors.initials: P
authors.lastname: Abraham
authors.affiliation: Department of Obstetrics and Gynaecology, Maulana Azad Medical College, New Delhi, India.
authors.forename: Reva
authors.initials: R
authors.lastname: Tripathi
authors.affiliation: Pediatric Biology Center, Translational Health Science and Technology institute, NCR Biotech Science Cluster, Faridabad, India.
authors.forename: Nitya
authors.initials: N
authors.lastname: Wadhwa
authors.affiliation: Pediatric Biology Center, Translational Health Science and Technology institute, NCR Biotech Science Cluster, Faridabad, India.
authors.forename: Shinjini
authors.initials: S
authors.lastname: Bhatnagar
authors.affiliation: Madras Diabetes Research Foundation, No. 4, Conran Smith Road, Gopalapuram, Chennai, India.
authors.forename: Visvanathan Gnana
authors.initials: VG
authors.lastname: Prakash
authors.affiliation: Madras Diabetes Research Foundation, No. 4, Conran Smith Road, Gopalapuram, Chennai, India.
authors.forename: Venkatesan
authors.initials: V
authors.lastname: Radha
authors.affiliation: Madras Diabetes Research Foundation, No. 4, Conran Smith Road, Gopalapuram, Chennai, India.
authors.forename: R M
authors.initials: RM
authors.lastname: Anjana
authors.affiliation: Madras Diabetes Research Foundation, No. 4, Conran Smith Road, Gopalapuram, Chennai, India.
authors.forename: V
authors.initials: V
authors.lastname: Mohan
authors.affiliation: Dept. of Microbiology and Immunology, Graduate School of Medicine, Osaka University, Japan.
authors.forename: Kiyoshi
authors.initials: K
authors.lastname: Takeda
authors.affiliation: Dept. of Microbiology and Immunology, Graduate School of Medicine, Osaka University, Japan.
authors.forename: Takashi
authors.initials: T
authors.lastname: Kurakawa
authors.affiliation: Molecular Genetics Laboratory, Centre for Human Microbial Ecology, Translational Health Science and Technology Institute, NCR Biotech Science Cluster, Faridabad, India.
authors.forename: G Balakrish
authors.initials: GB
authors.lastname: Nair
authors.affiliation: Molecular Genetics Laboratory, Centre for Human Microbial Ecology, Translational Health Science and Technology Institute, NCR Biotech Science Cluster, Faridabad, India.
authors.forename: Bhabatosh
authors.initials: B
authors.lastname: Das
mesh_terms: D004247:DNA / Q000302:isolation & purification*; D004269:DNA, Bacterial / Q000302:isolation & purification; D064349:Genome, Microbial; D059014:High-Throughput Nucleotide Sequencing; D006801:Humans; D056186:Metagenomics / Q000379:methods*; D012988:Soil Microbiology
chemical_list: D004269:DNA, Bacterial; D004247:DNA