No rule to produce assembly

[mhmism]

Hello,

I would like to run Hecatomb only using the assembly or ctg_annotations module, but I end up with an error.

Here is an example code: hecatomb run --reads /reads ctg_annotations --threads 16

I get this error: Building DAG of jobs... MissingRuleException: No rule to produce ctg_annotations (if you use input functions make sure that they don't raise unexpected exceptions).

[beardymcjohnface]

Hi,

The target is ctg_annotate. Try this:

hecatomb run --reads reads/ ctg_annotate --threads 16

[mhmism]

Thanks! You are right. I think it is working now. localrules: all, preprocess, assemble, annotate, ctg_annotate, print_stages, dumpSamplesTsv

I am closing the issue

[mhmism]

I have reopened the issue because I have a question related to the annotation. I am interested only in the ctg_annotate module (contigs preprocessing, assembly, and contigs annotations). If I am not mistaken, I think the reads sequence table is also being fed to the mmseqs2 annotation (even if I only run Hecatomb with the ctg_annotate rule). Is this correct? Is there any reason for this? If so, can I bypass the read sequence annotation and move directly to the contigs annotation by mmseqs2?

Here is the command output that comes from Hecatomb: { # Run mmseqs taxonomy module mmseqs easy-taxonomy hecatomb.out/results/seqtable.fasta /home/.conda/envs/hecatomb/lib/python3.10/site-packages/hecatomb/snakemake/workflow/../databases/aa/virus_primary_aa/sequenceDB hecatomb.out/processing/mmseqs_aa_primary/MMSEQS_AA_PRIMARY hecatomb.out/processing/mmseqs_aa_primary/mmseqs_aa_tmp --min-length 30 -e 1e-3 --start-sens 1 --sens-steps 3 -s 7 --lca-mode 2 --shuffle 0 -a --tax-output-mode 2 --search-type 2 --tax-lineage 1 --lca-ranks "superkingdom,phylum,class,order,family,genus,species" --format-output "query,target,evalue,pident,fident,nident,mismatch,qcov,tcov,qstart,qend,qlen,tstart,tend,tlen,alnlen,bits,qheader,theader,taxid,taxname,taxlineage" --threads 24 --split-memory-limit 48000M;

[beardymcjohnface]

Yes, it's dumb. The issue is that there are both direct annotations of the contigs and read-based annotations of the contigs in the targets for ctg_annotate, so it's essentially just another target for running everything. I'll fix it in the next version. Until then, I'd suggest just running hecatomb run assemble ... and then manually annotate with mmseqs using the hecatomb secondary nt database.

[mhmism]

Hello, thanks a lot. It will be great if this can be modified in the next version.

In the meantime, here is the code that I retrieved from a previous Hecatom run to make the annotations for the contigs manually (in case anyone is curious about it):

Create directories for the output tmp files

mkdir home/test/hecatomb.out/results/queryDB mkdir home/test/hecatomb.out/results/mmseqs2_results mkdir home/test/hecatomb.out/results/tophit

Create an mmseqs2 query database

mmseqs createdb home/test/hecatomb.out/results/cross_assembly.fasta home/test/hecatomb.out/results/queryDB/queryDB --dbtype 2;

Make an mmseqs2 search against the Hecatomb secondary nt database

mmseqs search home/test/hecatomb.out/results/queryDB/queryDB /home/.conda/envs/hecatomb/lib/python3.10/site-packages/hecatomb/snakemake/databases/nt/virus_secondary_nt/sequenceDB home/test/hecatomb.out/results/mmseqs2_results/mmseqs2_results home/test/hecatomb.out/results/mmseqs2_tmp --start-sens 2 -s 7 --sens-steps 3 --split-memory-limit 22000M --min-length 90 -e 1e-20 --search-type 3 --threads 16 > home/test/hecatomb.out/results/mmseqs_contig_annotation.log

Filter TopHit results

mmseqs filterdb home/test/hecatomb.out/results/mmseqs2_results/mmseqs2_results home/test/hecatomb.out/results/tophit/tophit --extract-lines 1;

Convert to alignments

mmseqs convertalis home/test/hecatomb.out/results/queryDB/queryDB /home/.conda/envs/hecatomb/lib/python3.10/site-packages/hecatomb/snakemake/databases/nt/virus_secondary_nt/sequenceDB home/test/hecatomb.out/results/tophit/tophit home/test/hecatomb.out/results/tophit/tophit.m8 --format-output 'query,target,evalue,pident,fident,nident,mismatch,qcov,tcov,qstart,qend,qlen,tstart,tend,tlen,alnlen,bits,target';

Header for output table

printf "contigID evalue pident fident nident mismatch qcov tcov qstart qend qlen tstart tend tlen alnlen bits target kingdom phylum class order family genus species" > home/test/hecatomb.out/results/contigAnnotations.tsv;

Assign taxonomy

sed 's/tid|//' home/test/hecatomb.out/results/tophit/tophit.m8 | sed 's/|\S*//' | taxonkit lineage --data-dir /home/.conda/envs/hecatomb/lib/python3.10/site-packages/hecatomb/snakemake/databases/tax/taxonomy -i 2 | taxonkit reformat --data-dir /home/.conda/envs/hecatomb/lib/python3.10/site-packages/hecatomb/snakemake/databases/tax/taxonomy -i 19 -f '{k}\t{p}\t{c}\t{o}\t{f}\t{g}\t{s}' -F --fill-miss-rank | cut --complement -f2,19 >> home/test/hecatomb.out/results/contigAnnotations.tsv;