Bio Binning QC

Bin metagenomic contigs, route bins by domain, run domain-specific quality checks, and join the results into normalized tables.

Instructions

Tool guides and versions: docs/README.md.

1. Map each sample's reads back to the assembly (BBMap or bwa-mem2 for short reads, minimap2 for long reads; see /bio-reads-qc-mapping), then compute per-sample contig depth from the sorted BAMs with CoverM v0.7.0+.
2. Bin contigs with QuickBin from Bryce Foster's BBTools container (bryce911/bbtools:39.85; record the digest when pulled). QuickBin is the default for short-read and long-read assemblies (docs/quickbin.md). On a CUDA GPU node, SemiBin2 v2.3.0+ is the alternative. Use MetaBAT2 v2.18+ only to reproduce a prior pipeline.
3. Run /tracking-taxonomy-updates for BBTools-container QuickClade domain triage on the bin directory and the source assembly with percontig. Persist the per-contig screen so mixed bins are visible.
4. Route bins by the QuickClade domain screen:

• Bacteria or Archaea -> run GTDB-Tk taxonomy assignment. If the GTDB-Tk reference package is missing, set it up under $BIO_DB_ROOT, export GTDBTK_DATA_PATH, run gtdbtk check_install, and record the release before classification.
• Eukaryota -> run EukCC v2.1.3+ for eukaryotic bins.
• Viral or virus-like -> remove from MAG QC and route candidate contigs/genomes to /bio-viromics; use vConTACT3 for phage/prokaryotic-virus evidence and GVClass for giant-virus/Nucleocytoviricota candidates.
• Mixed or low-confidence -> flag as potential chimeras and inspect per-contig assignments before QC scoring.

1. Run domain-specific QC:

• CheckM2 v1.1.0+ for bacterial and archaeal bins (v1.1.0 is a breaking upgrade: update the pinned Pixi environment and refresh the DIAMOND v3 database from Zenodo DOI 10.5281/zenodo.14897628).
• EukCC v2.1.3+ for eukaryotic bins.
• GUNC v1.1.0+ (needed for the progenomes_3 and gtdb_214 databases) for bacterial and archaeal bins only, as a chimerism check next to CheckM2. Do not apply GUNC to eukaryotic bins. gunc run reads .fa files from --input_dir by default; set --file_suffix to match other bin suffixes.

1. Normalize routed outputs with scripts/build_bin_qc_tables.py. The join rejects GUNC rows for non-prokaryotic routes, refuses prokaryotic or eukaryotic bins that lack their domain-specific QC or taxonomy outputs, and refuses to overwrite existing outputs. Accepted routes are prokaryote, eukaryote, prokaryotic_virus, giant_virus, and manual_review.

Quick Reference

Input Requirements

Prerequisites:

• Tools declared in the project's pinned Pixi environment. See docs/README.md for expected tools.
• Reference DB root: set BIO_DB_ROOT to the project or site-local database directory.
• Coverage/depth tables or reads available to compute coverage.
• Docker or Apptainer/Singularity available for bryce911/bbtools QuickBin runs, or a documented local BBTools install.

Inputs:

• contigs.fasta
• coverage.tsv (per-sample depth table)

Output

• results/bio-binning-qc/bins/ (QuickBin or SemiBin2 output)
• results/bio-binning-qc/quickclade_percontig.tsv and domain_routing.tsv (from /tracking-taxonomy-updates)
• results/bio-binning-qc/bin_metrics.tsv (one row per routed bin; written by build_bin_qc_tables.py)
• results/bio-binning-qc/gtdbtk_taxonomy.tsv (one row per prokaryotic bin; written by build_bin_qc_tables.py)
• results/bio-binning-qc/bin_qc_report.json (routed-bin counts by domain; written by build_bin_qc_tables.py)
• results/bio-binning-qc/logs/

Quality Gates

• [ ] Completeness, contamination, and GUNC chimerism results are reported against project thresholds (for example MIMAG, see docs/README.md).
• [ ] On execution failure, preserve logs and report the failed command; retry only after diagnosing the cause and recording the changed parameters. Report unmet biological thresholds as results; never tune parameters solely to pass a gate.
• [ ] Verify contigs.fasta and coverage.tsv are non-empty.
• [ ] Verify reference DBs for QC tools exist under the reference root.
• [ ] QuickClade percontig screen exists for the source assembly and bin set before CheckM2/EukCC/GTDB-Tk decisions.
• [ ] Bacterial and archaeal bins have GTDB-Tk taxonomy with the database release recorded.
• [ ] Viral/virus-like bins are routed to /bio-viromics instead of reported as MAGs.
• [ ] Mixed-domain bins are flagged as possible contamination/chimeras with per-contig evidence.
• [ ] bin_metrics.tsv covers every routed bin, including viral and manual-review rows, and gtdbtk_taxonomy.tsv covers every prokaryotic bin.