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. 2024 Apr 29;14:9785. doi: 10.1038/s41598-024-57981-4

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© This is a U.S. Government work and not under copyright protection in the US; foreign copyright protection may apply 2024

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/.

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For the DNA mixtures, independently measured ‘ground truth’ results (black 99% confidence intervals) for the ratios between taxa relative abundances can be compared to each individual lab’s amplicon (red points) or shotgun (blue points) metagenomic sequencing results, as well as the range of results (grey boxplots) among participating labs. The taxa in Mix A were roughly equally abundant, while the Mix B sample exhibited groups of taxa added at tenfold dilutions. The horizontal axis identifies the taxa (known to be present in the DNA mixtures) whose observed relative abundances were ratioed. The ground truth values were scaled to account for known 16S copy numbers (for amplicon sequencing) or genome sizes (for shotgun sequencing), so the ‘actual’ ratios vary slightly between the two analyses even though the DNA concentrations are identical. Genus-level taxonomic bar charts by (16S and WGS analysis) show the average composition observed for each DNA mixture (Fig. SI-10).