Frontiers in Microbiology · 2019 · 119 citations · 49 references
BiologyHuman BodyUnicellular OrganismConventional Microbiome SurveysMicrobial SystematicsMicrobial DiversityBiosignatureExtremophileNasal CavityArchaeal SignaturesMicrobial EcologyMicrobiologyArchaeaMicrobiomeMedicineBioinformaticsHigh Throughput SequencingAmplicon Sequencing
Due to their fundamentally different biology, archaea are consistently overlooked in conventional microbiome surveys. Using amplicon sequencing, we evaluated methodological set-ups to detect archaea in samples from five different body sites: respiratory tract (nasal cavity), digestive tract (mouth, appendix, and stool) and skin. With optimized protocols, the detection of archaeal ribosomal sequence variants (RSVs) was increased from one (found in currently used, so-called "universal" approach) to 81 RSVs in a representative sample set. The results from this extensive primer-evaluation led to the identification of the primer pair combination 344f-1041R/519F-806R which performed superior for the analysis of the archaeome of gastrointestinal tract, oral cavity and skin. The proposed protocol might not only prove useful for analyzing the human archaeome in more detail but could also be used for other holobiont samples.
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DADA2: High-resolution sample inference from Illumina amplicon data
Benjamin J. Callahan, Paul J. McMurdie, Michael Rosen et al. · Nature Methods · 2016 · 33.5K citations · Full text
Illumina Amplicon Data, Omics Datasets, Spectral Searching +5
The SILVA ribosomal RNA gene database project: improved data processing and web-based tools
Christian Quast, Elmar Pruesse, Pelin Yilmaz et al. · Nucleic Acids Research · 2012 · 32.2K citations · Full text
Ribosomal Rna, Genetics, Genomics +20