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Topics {✒️}

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Questions {❓}

• Branton WG, Ellestad KK, Maingat F, Wheatley BM, Rud E, Warren RL, Holt RA, Surette MG, Power C: Brain microbial populations in HIV/AIDS: ?

Schema {🗺️}

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      headline:Reagent and laboratory contamination can critically impact sequence-based microbiome analyses
      description:The study of microbial communities has been revolutionised in recent years by the widespread adoption of culture independent analytical techniques such as 16S rRNA gene sequencing and metagenomics. One potential confounder of these sequence-based approaches is the presence of contamination in DNA extraction kits and other laboratory reagents. In this study we demonstrate that contaminating DNA is ubiquitous in commonly used DNA extraction kits and other laboratory reagents, varies greatly in composition between different kits and kit batches, and that this contamination critically impacts results obtained from samples containing a low microbial biomass. Contamination impacts both PCR-based 16S rRNA gene surveys and shotgun metagenomics. We provide an extensive list of potential contaminating genera, and guidelines on how to mitigate the effects of contamination. These results suggest that caution should be advised when applying sequence-based techniques to the study of microbiota present in low biomass environments. Concurrent sequencing of negative control samples is strongly advised.
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      name:Molecular Genetics and Genomics, National Heart and Lung Institute, Imperial College London, London, UK
      name:Molecular Genetics and Genomics, National Heart and Lung Institute, Imperial College London, London, UK
      name:Shoklo Malaria Research Unit, Mahidol-Oxford Tropical Medicine Research Unit, Faculty of Tropical Medicine, Mahidol University, Mae Sot, Thailand
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      name:Pathogen Genomics Group, Wellcome Trust Sanger Institute, Hinxton, UK
      name:Institute of Microbiology and Infection, University of Birmingham, Birmingham, UK
      name:Pathogen Genomics Group, Wellcome Trust Sanger Institute, Hinxton, UK
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