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dc.contributor.authorSusannah J. Salteren_US
dc.contributor.authorMichael J. Coxen_US
dc.contributor.authorElena M. Tureken_US
dc.contributor.authorSzymon T. Calusen_US
dc.contributor.authorWilliam O. Cooksonen_US
dc.contributor.authorMiriam F. Moffatten_US
dc.contributor.authorPaul Turneren_US
dc.contributor.authorJulian Parkhillen_US
dc.contributor.authorNicholas J. Lomanen_US
dc.contributor.authorAlan W. Walkeren_US
dc.contributor.otherWellcome Trust Sanger Instituteen_US
dc.contributor.otherNational Heart and Lung Instituteen_US
dc.contributor.otherUniversity of Birminghamen_US
dc.contributor.otherMahidol Universityen_US
dc.contributor.otherNuffield Department of Clinical Medicineen_US
dc.contributor.otherUniversity of Aberdeenen_US
dc.identifier.citationBMC Biology. Vol.12, No.1 (2014)en_US
dc.description.abstract© 2014 Salter et al. Background: 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. Results: 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. Conclusions: 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.en_US
dc.rightsMahidol Universityen_US
dc.subjectAgricultural and Biological Sciencesen_US
dc.subjectBiochemistry, Genetics and Molecular Biologyen_US
dc.titleReagent and laboratory contamination can critically impact sequence-based microbiome analysesen_US
Appears in Collections:Scopus 2011-2015

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