Phyloseq objects used in the workflow (ps_*)

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ps_* object What it contains Taxonomic composition Alpha diversity Beta diversity DESeq2 differential abundance
ps_raw Raw imported phyloseq object (integer counts; as imported) ❌ (not recommended directly) ⚠️ only if you also apply sample filtering first
ps_base ps_raw + taxonomy + sample metadata aligned (still raw counts) ⚠️ only if you also apply sample filtering first
ps_pruned Optional subset of ps_base (e.g., remove unwanted samples by ID/pattern); still raw counts ⚠️ only if you also apply low-depth filtering
ps_filt Filtered samples (low-depth samples removed) + taxa with nonzero totals; absolute counts ✅ as a starting point (but plot on ps_rel) ✅ as the input to rarefaction ✅ as the input to rarefaction ✅ as the input to ps_deseq
ps_rel Relative abundance (compositional) computed from ps_filt ✅ primary
ps_abund Absolute counts after “plotting taxa filter” (e.g., mean rel. abundance > 0.1%), derived from ps_filt via ps_rel ✅ (if you want cleaner plots) ❌ (not recommended)
ps_abund_rel Relative abundance computed from ps_abund (filtered taxa set) ✅ (clean composition plots)
ps_rarefied Rarefied counts from ps_filt (even depth) ✅ primary ✅ primary
ps_deseq Non-rarefied integer counts from ps_filt + optional count-based taxon prefilter (e.g., total ≥ 10) ✅ primary

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