Effect of storage and DNA extraction method on 16S rRNA-profiled fecal microbiota in Japanese adults
作者： Yuki Kawada , Yuji Naito , Akira Andoh , Motoyuki Ozeki , Ryo Inoue
DOI: 10.3164/JCBN.18-84
关键词: 16S ribosomal RNA 、 DNA 、 DNA extraction 、 Bacteroides 、 Bifidobacterium 、 Enzyme 、 Feces 、 Food science 、 Ethanol precipitation 、 Chemistry
摘要: The effect of two factors, storage and the bacterial DNA extraction method, that potentially affect 16S rRNA-based profiling microbiota in feces Japanese adults, were evaluated. Profiles stored DESS (DMSO-EDTA-salt solution) for 1, 2 3 weeks at room temperature, 4°C compared with those fresh guanidine thiocyanate solution 4°C. None variables (preservation solution, temperature duration) considerably affected α- β-diversity fecal OTU profiles. Regarding methods, four evaluated; A) silica membrane purification combined bead-beating disruption, B) magnetic bead C) manual using phenol-chloroform ethanol precipitation enzymatic lysis, D) by a commercially available stool kit. While methods A, B, C did not markedly profiles, method D noticeably altered both β-diversity. In addition, caused significant changes abundance predominant genera; Bacteroides Bifidobacterium.
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