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

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作者
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
机构
[1] Molecular Genetics Laboratory,Department of Gastroenterology
[2] Centre for Human Microbial Ecology,Department of Obstetrics and Gynaecology
[3] Translational Health Science and Technology Institute,undefined
[4] NCR Biotech Science Cluster,undefined
[5] Section of Metabolic Genetics,undefined
[6] Novo Nordisk Foundation Center for Basic Metabolic Research,undefined
[7] University of Copenhagen,undefined
[8] P.D. Hinduja Hospital and Medical Research Centre,undefined
[9] Maulana Azad Medical College,undefined
[10] Pediatric Biology Center,undefined
[11] Translational Health Science and Technology institute,undefined
[12] NCR Biotech Science Cluster,undefined
[13] Madras Diabetes Research Foundation,undefined
[14] Dept. of Microbiology and Immunology,undefined
[15] Graduate School of Medicine,undefined
[16] Osaka University,undefined
来源
Scientific Reports | / 6卷
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摘要
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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