Quantifying variation across 16S rRNA gene sequencing runs in human microbiome studies

• DOI: 10.1007/s00253-024-13198-z
• Publication Date: 2024-06-08T10:01:23Z
• Authors: Andrew J. Hoisington; Christopher E. St...; Joseph C. Ellis; Christopher A. Lowry; Lisa A. Brenner
• Keywords: Variation (astronomy)

Abstract Recent microbiome research has incorporated a higher number of samples through more participants in study, longitudinal studies, and metanalysis between studies. Physical limitations sequencing machine can result spread across runs. Here we present the results nearly 1000 16S rRNA gene sequences fecal (stabilized swab) oral (swab) from multiple human studies positive controls that were conducted with identical standard operating procedures. Sequencing was performed same center 18 different The simplified mock community showed accuracy, while precision (e.g., technical variation) robust for actual control samples. Technical variation lowest stabilized samples, followed by swab then order stability inverse DNA concentrations highest samples), highlighting importance concentration reproducibility urging caution when analyzing low biomass Coefficients at genus level also trend lower concentrations. both sample types two sampling locations significantly less than observed biological variation. Overall, this providing comparisons variation, highlights using controls, provides semi-quantified data to better understand introduced Key points • Mock accuracy precision. Samples had increased Biological due