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. 2022 Feb 22;13:863. doi: 10.1038/s41467-022-28426-1

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© The Author(s) 2022

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 license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license 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 license, visit http://creativecommons.org/licenses/by/4.0/.

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Fig. 2 — a Comparison of the 16S rRNA sequences identified by Sanger sequencing and by BarBIQ. Edit distance, Levenshtein distance⁶⁸, defined as the minimum number of substitutions, insertions, and deletions; San sequence, Sanger sequencing-identified 16S rRNA sequence; ATCC/JCM/DSM <number>, strain ID; A, B, or C, San sequences for each strain; Bar-sequence-MK-<number>, BarBIQ-identified sequences (Bar sequences); COTU-MK-<number>, cell-based operational taxonomy units (cOTUs); red star, Bar sequences that had one base difference. Source data are provided as a Source Data file. b Comparison (Venn diagram) among San sequences identified from each of the ten strains, Bar sequences, amplicon sequence variants (ASVs), and the representative sequences of operational taxonomy units (OTU-RepSeqs) identified from the mock community. Circle, total sequences from each method; numbers in circles, number of unique or identical sequences detected by the given method(s); numbers in the parentheses, total number of sequences detected by the given method. c Comparison of the absolute cell abundance per unit volume of 10 strains in the mock community measured by BarBIQ ([C]_BarBIQ) (Supplementary Data 6) and by microscopic imaging ([C]_Microscope) (Supplementary Table 1). Data are presented as mean values ± SD (n = 3 for [C]_BarBIQ, n = 5 for [C]_Microscope). Blue thin line, a fitting line with a fixed slope of one in log scale (intercept: -0.035) by considering the standard errors of both [C]_BarBIQ and [C]_Microscope, indicating the averaged ratio [C]_BarBIQ/[C]_Microscope as 0.92; gray thick line, 95% confidence interval of the fitted line, indicating the 95% confidence interval of the ratio [C]_BarBIQ/[C]_Microscope as 0.68~1.25; r, Pearson coefficient; R², coefficient of determination. d Comparison of the proportional abundances of 15 ASVs in the mock community measured by Conv_ASV (proportional [C]_ASV) (Supplementary Data 5) with the proportional [C]_Microscope measured by microscopic imaging. Data are presented as mean values ± SD (n = 2 for proportional [C]_ASV, n = 5 for proportional [C]_Microscope). Strains that had commonly detected sequence(s) are shown. The strains that were compared with multiple identical ASVs are shown in colors. By the same fitting as c (intercept: −0.28), the averaged ratio of proportional [C]_ASV and proportional [C]_Microscope was 0.52, and its 95% confidence interval was 0.28–0.99.