The UniFrac distance metric is often used to separate groups in microbiome analysis, but Food Storage Containers requires a constant sequencing depth to work properly.Here we demonstrate that unweighted UniFrac is highly sensitive to rarefaction instance and to sequencing depth in uniform data sets with no clear structure or separation between groups.We show that this arises because of subcompositional effects.
We introduce information UniFrac and ratio UniFrac, two new weightings that are not as sensitive to rarefaction and allow greater separation of outliers than classic unweighted and loader weighted UniFrac.With this expansion of the UniFrac toolbox, we hope to empower researchers to extract more varied information from their data.