#!/bin/bash set -e # ============================================================================= # Task 27: Read Downsampling and Assembly Quality Titration # # DAG (depth 5, replicated fan-out): # # L0: PE reads + reference # L1: fastp (trim full dataset) # L2: ├── seqkit sample 25% → megahit → quast # ├── seqkit sample 50% → megahit → quast # └── seqkit sample 100% → megahit → quast # L3: Parse all three QUAST reports # L4: MERGE (titration report: how coverage affects assembly) # ============================================================================= 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" REFERENCE="${REF}/reference.fna" FRACTIONS=("0.25" "0.50" "1.00") log_step() { echo "==================================================================" echo "STEP: $1" echo "$(date)" echo "==================================================================" } mkdir -p "${OUT}/trimmed" "${RES}" # L1: Trim log_step "L1: fastp" if [ ! -f "${OUT}/trimmed/R1.fastq.gz" ]; then fastp --in1 "${DATA}/reads_R1.fastq.gz" --in2 "${DATA}/reads_R2.fastq.gz" \ --out1 "${OUT}/trimmed/R1.fastq.gz" --out2 "${OUT}/trimmed/R2.fastq.gz" \ --detect_adapter_for_pe --thread ${THREADS} --json "${OUT}/trimmed/fastp.json" else echo "Skipping (exists)"; fi # L2: For each fraction, subsample + assemble + QC for FRAC in "${FRACTIONS[@]}"; do FRAC_DIR="${OUT}/frac_${FRAC}" mkdir -p "${FRAC_DIR}" log_step "L2: Subsample ${FRAC} + assemble" if [ ! -f "${FRAC_DIR}/assembly/final.contigs.fa" ]; then if [ "${FRAC}" = "1.00" ]; then cp "${OUT}/trimmed/R1.fastq.gz" "${FRAC_DIR}/R1.fastq.gz" cp "${OUT}/trimmed/R2.fastq.gz" "${FRAC_DIR}/R2.fastq.gz" else seqkit sample -p "${FRAC}" -s 42 "${OUT}/trimmed/R1.fastq.gz" -o "${FRAC_DIR}/R1.fastq.gz" seqkit sample -p "${FRAC}" -s 42 "${OUT}/trimmed/R2.fastq.gz" -o "${FRAC_DIR}/R2.fastq.gz" fi megahit -1 "${FRAC_DIR}/R1.fastq.gz" -2 "${FRAC_DIR}/R2.fastq.gz" \ -o "${FRAC_DIR}/assembly" -t ${THREADS} --min-contig-len 500 else echo "Skipping (exists)"; fi log_step "L2: QUAST ${FRAC}" if [ ! -f "${FRAC_DIR}/quast/report.tsv" ]; then quast "${FRAC_DIR}/assembly/final.contigs.fa" -r "${REFERENCE}" \ -o "${FRAC_DIR}/quast" -t ${THREADS} else echo "Skipping (exists)"; fi done # L3-L4: Parse + MERGE log_step "MERGE" echo "fraction,total_length,num_contigs,n50,largest_contig,genome_fraction,num_reads" > "${RES}/downsampling_report.csv" for FRAC in "${FRACTIONS[@]}"; do FRAC_DIR="${OUT}/frac_${FRAC}" TOTAL=$(grep "^Total length" "${FRAC_DIR}/quast/report.tsv" | head -1 | cut -f2) NCTG=$(grep "^# contigs " "${FRAC_DIR}/quast/report.tsv" | head -1 | cut -f2) N50=$(grep "^N50" "${FRAC_DIR}/quast/report.tsv" | cut -f2) LARGEST=$(grep "^Largest contig" "${FRAC_DIR}/quast/report.tsv" | cut -f2) GF=$(grep "^Genome fraction" "${FRAC_DIR}/quast/report.tsv" | cut -f2) NREADS=$(zcat "${FRAC_DIR}/R1.fastq.gz" | wc -l | awk '{print $1/4}') echo "${FRAC},${TOTAL},${NCTG},${N50},${LARGEST},${GF},${NREADS}" >> "${RES}/downsampling_report.csv" done echo "" echo "=== Pipeline complete ===" cat "${RES}/downsampling_report.csv"