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Box plots of Euclidean distances between observed and expected relative species abundances support accurate assessment of true bacterial mock community composition. Euclidean distances between observed and expected relative species abundances of a mock bacterial community were calculated for four technical replicates of the V1–V3 and V3–V4 regions using OTUs picked with the closed-reference protocol against Greengenes (GG) and HOMD. The x-axis represents the different amplicon/OTU picking method used, and the y-axis represents the distance from expected values