期刊
SYSTEMATIC AND APPLIED MICROBIOLOGY
卷 41, 期 2, 页码 151-157出版社
ELSEVIER GMBH, URBAN & FISCHER VERLAG
DOI: 10.1016/j.syapm.2017.11.008
关键词
Human fecal sample; Gut microbiota; DNA extraction; Bead beating; 16S rRNA gene sequencing
资金
- Main Research Program of the Korea Food Research Institute (KFRI) - Ministry of Science and ICT [E0170602-01]
The human gut harbors a vast range of microbes that have significant impact on health and disease. Therefore, gut microbiome profiling holds promise for use in early diagnosis and precision medicine development. Accurate profiling of the highly complex gut microbiome requires DNA extraction methods that provide sufficient coverage of the original community as well as adequate quality and quantity. We tested nine different DNA extraction methods using three commercial kits (TianLong Stool DNA/RNA Extraction Kit (TS), QIAamp DNA Stool Mini Kit (QS), and QIAamp PowerFecal DNA Kit (QP)) with or without additional bead-beating step using manual or automated methods and compared them in terms of DNA extraction ability from human fecal sample. All methods produced DNA in sufficient concentration and quality for use in sequencing, and the samples were clustered according to the DNA extraction method. Inclusion of bead-beating step especially resulted in higher degrees of microbial diversity and had the greatest effect on gut microbiome composition. Among the samples subjected to bead-beating method, TS kit samples were more similar to QP kit samples than QS kit samples. Our results emphasize the importance of mechanical disruption step for a more comprehensive profiling of the human gut microbiome. (C) 2017 The Authors. Published by Elsevier GmbH.
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