Pipeline sensitivity and assessment of the relative abundance accuracy.

<p><b>(a)</b> Stacked bar charts showing the relative abundance of bacterial families obtained by sequencing of a serial dilution of the plasmid-based mock community (from 10 and 10⁷ copies of 16S gene for each of the 14 bacterial strains) compared to the actual relative abundance (right stacked bar). <b>(b)</b> Generalized UniFrac distance based PCoA analysis of the sequencing of a serial dilution of the plasmid-based mock community shown in a compared with the negative control generated by sequencing molecular biology-grade water with the same pipeline (H₂O). UniFrac weight parameter (Alpha) was set to 0.6 for this analysis. <b>(c)</b> Stacked bar charts showing the relative abundance of bacterial families obtained by sequencing 7 mixes with different ratios of two of the bacterial plasmids containing the 16S gene of <i>Geofilum rubicundum</i> (GR) and <i>Eubacterium barkeri</i> (EB). The sequencing was performed in triplicate for all the samples (starting from the extracted DNA); the means of the triplicates are shown in <b>a</b> and individual triplicates are shown in <b>b</b> and <b>c</b>.