Optimisation of methods for bacterial skin microbiome investigation: primer selection and comparison of the 454 versus MiSeq platform

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• 作者：Madhura Castelino ; Stephen Eyre ; John Moat ; Graeme Fox ; Paul Martin…
• 关键词：Skin microbiome ; Bacterial microbiome ; Low biomass ; 16s rRNA gene ; Miseq platform ; Next generation sequencing ; Methods ; Optimisation ; Primer selection
• 刊名：BMC Microbiology
• 出版年：2017
• 出版时间：December 2017
• 年：2017
• 卷：17
• 期：1
• 全文大小：1382KB
• 刊物主题：Microbiology; Biological Microscopy; Mycology; Parasitology; Virology; Life Sciences, general;
• 出版者：BioMed Central
• ISSN：1471-2180
• 卷排序：17

文摘

BackgroundThe composition of the skin microbiome is predicted to play a role in the development of conditions such as atopic eczema and psoriasis. 16S rRNA gene sequencing allows the investigation of bacterial microbiota. A significant challenge in this field is development of cost effective high throughput methodologies for the robust interrogation of the skin microbiota, where biomass is low. Here we describe validation of methodologies for 16S rRNA (ribosomal ribonucleic acid) gene sequencing from the skin microbiome, using the Illumina MiSeq platform, the selection of primer to amplify regions for sequencing and we compare results with the current standard protocols..