#!/bin/bash set -e # ============================================================================= # Task 29: Genome Completeness and Contamination Assessment # # DAG (depth 5): # L0: assembled genome # L1: ├── BUSCO (single-copy orthologs) # ├── QUAST (assembly metrics) # └── Prokka (gene prediction for stats) # L2: ├── seqkit stats (basic stats) # └── parse BUSCO/QUAST/Prokka outputs # L3: checkv (if viral) OR manual assessment # L4: MERGE (comprehensive quality report) # ============================================================================= THREADS=$(( $(nproc) > 8 ? 8 : $(nproc) )) SCRIPT_DIR="$(cd "$(dirname "${BASH_SOURCE[0]}")" && pwd)" DATA="${SCRIPT_DIR}/data" REF="${SCRIPT_DIR}/reference" OUT="${SCRIPT_DIR}/outputs" RES="${SCRIPT_DIR}/results" GENOME="${DATA}/genome.fna" REFERENCE="${REF}/reference.fna" log_step() { echo "==================================================================" echo "STEP: $1" echo "$(date)" echo "==================================================================" } mkdir -p "${OUT}"/{busco_out,quast,prokka,stats} "${RES}" # L1-A: BUSCO log_step "L1-A: BUSCO" if [ ! -d "${OUT}/busco_out/busco" ]; then busco -i "${GENOME}" -o busco --out_path "${OUT}/busco_out" \ -l bacteria_odb10 -m genome -c ${THREADS} --force else echo "Skipping (exists)"; fi # L1-B: QUAST with reference log_step "L1-B: QUAST" if [ ! -f "${OUT}/quast/report.tsv" ]; then quast "${GENOME}" -r "${REFERENCE}" -o "${OUT}/quast" -t ${THREADS} else echo "Skipping (exists)"; fi # L1-C: Prokka log_step "L1-C: Prokka" if [ ! -f "${OUT}/prokka/genome.gff" ]; then prokka "${GENOME}" --outdir "${OUT}/prokka" --prefix genome \ --cpus ${THREADS} --kingdom Bacteria --force else echo "Skipping (exists)"; fi # L2: seqkit + parse log_step "L2: Parse results" seqkit stats "${GENOME}" -T > "${OUT}/stats/seqkit.tsv" # BUSCO BUSCO_SUM=$(grep "C:" "${OUT}/busco_out/busco/short_summary.specific.bacteria_odb10.busco.txt" 2>/dev/null \ | head -1 | sed 's/^[[:space:]]*//' || echo "N/A") BUSCO_COMPLETE=$(echo "$BUSCO_SUM" | grep -oP 'C:[\d.]+' | tr -d 'C:') BUSCO_FRAGMENTED=$(echo "$BUSCO_SUM" | grep -oP 'F:[\d.]+' | tr -d 'F:') BUSCO_MISSING=$(echo "$BUSCO_SUM" | grep -oP 'M:[\d.]+' | tr -d 'M:') # QUAST TOTAL_LEN=$(grep "^Total length" "${OUT}/quast/report.tsv" | head -1 | cut -f2) NUM_CONTIGS=$(grep "^# contigs " "${OUT}/quast/report.tsv" | head -1 | cut -f2) N50=$(grep "^N50" "${OUT}/quast/report.tsv" | cut -f2) GC=$(grep "^GC" "${OUT}/quast/report.tsv" | cut -f2) LARGEST=$(grep "^Largest contig" "${OUT}/quast/report.tsv" | cut -f2) GF=$(grep "^Genome fraction" "${OUT}/quast/report.tsv" | cut -f2) MISASSEMBLIES=$(grep "^# misassemblies" "${OUT}/quast/report.tsv" | cut -f2) DUPLICATION=$(grep "^Duplication ratio" "${OUT}/quast/report.tsv" | cut -f2) # Prokka CDS=$(grep "^CDS" "${OUT}/prokka/genome.txt" | awk '{print $2}') TRNA=$(grep "^tRNA" "${OUT}/prokka/genome.txt" | awk '{print $2}') RRNA=$(grep "^rRNA" "${OUT}/prokka/genome.txt" | awk '{print $2}') CODING_DENSITY=$(python3 -c " cds_len = 0 for line in open('${OUT}/prokka/genome.gff'): if '\tCDS\t' in line: parts = line.split('\t') cds_len += int(parts[4]) - int(parts[3]) + 1 print(f'{cds_len/${TOTAL_LEN}*100:.1f}') " 2>/dev/null || echo "N/A") # L4: MERGE log_step "L4-MERGE" cat > "${RES}/completeness_report.csv" << CSVEOF metric,value total_length,${TOTAL_LEN} num_contigs,${NUM_CONTIGS} n50,${N50} gc_content,${GC} largest_contig,${LARGEST} genome_fraction,${GF} misassemblies,${MISASSEMBLIES} duplication_ratio,${DUPLICATION} busco_complete_pct,${BUSCO_COMPLETE} busco_fragmented_pct,${BUSCO_FRAGMENTED} busco_missing_pct,${BUSCO_MISSING} cds_count,${CDS} trna_count,${TRNA} rrna_count,${RRNA} coding_density_pct,${CODING_DENSITY} CSVEOF echo "" echo "=== Pipeline complete ===" cat "${RES}/completeness_report.csv"