Hi!

I run picrust2.5.1 using this code:

place_seqs.py -s seq.fa -o out.tre -p 30 --intermediate intermediate/place_seqs

hsp.py -i 16S -t out.tre -o marker_predicted_and_nsti.tsv.gz -p 30 -n

hsp.py -i EC -t out.tre -o EC_predicted.tsv.gz -p 30

metagenome_pipeline.py -i gr.biom -m marker_predicted_and_nsti.tsv.gz -f EC_predicted.tsv.gz -o EC_metagenome_out --strat_out

pathway_pipeline.py -i EC_metagenome_out/pred_metagenome_contrib.tsv.gz -o pathways_out -p 1

After the analysis I got this warning:

/soft/picrust2/picrust2-2.5.1/picrust2/pathway_pipeline.py:642: FutureWarning: The operation <function sum at 0x1489d7756160> failed on a column. If any error is raised, this will raise an exception in a future version of pandas. Drop these columns to avoid this warning.

Finally I got the unstratified and the stratified output, but I am bot sure if I can use it. Is it valid?

I also want to ask about the stratified table with predicted MetaCyc pathways. In the tutorial it is said that pathways table is similar to the EC table. However, the table with predicted pathways lacks the last column "norm_taxon_function_contrib" which gives proportional contribution of EC . Is this because such information is calculated only if I use --per_sequence_contrib option? If I use this option and I get the stratified table with pathways per sequence, is the last column also the proportional contribution of the given pathways? Is this last column analogous to the norm_taxon_function_contribution? Could you please clarify this?

Many thanks,
Kinga