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#!/bin/bash
#SBATCH --cpus-per-task=1
#SBATCH --mem=16GB
#SBATCH --time=24:00:00
#SBATCH --job-name=process_trio
#SBATCH --output=process_solo.%A_%a.out
#SBATCH --error=process_solo.%A_%a.err
# setup PATH
export PATH=$PATH:/home/u035/u035/shared/software/bcbio/anaconda/envs/python2/bin:/home/u035/u035/shared/software/bcbio/anaconda/bin
export PERL5LIB=$PERL5LIB:/home/u035/u035/shared/software/bcbio/anaconda/lib/site_perl/5.26.2
### folder structure for the downstream analysis - created by trio_setup.sh ###
BASE=/home/u035/u035/shared/analysis/work
WORK_DIR=$BASE/${PROJECT_ID}
VCF_DIR=${WORK_DIR}/VCF
PED_DIR=${WORK_DIR}/PED
LOG_DIR=${WORK_DIR}/LOG
G2P_DIR=${WORK_DIR}/G2P
VASE_DIR=${WORK_DIR}/VASE
COV_DIR=${WORK_DIR}/COV
DEC_DIR=${WORK_DIR}/DECIPHER
IGV_DIR=${DEC_DIR}/IGV
CNV_DIR=${WORK_DIR}/CNV
BAMOUT_DIR=${WORK_DIR}/BAMOUT
SCRIPTS_DIR=/home/u035/u035/shared/scripts
# other files to be used
FAMILY_IDS=${WORK_DIR}/solo_FAM_IDs.txt # created by trio_setup.sh
CHILD_IDS=${WORK_DIR}/solo_PRO_IDs.txt # created by trio_setup.sh
TARGETS=/home/u035/u035/shared/resources/G2P/DDG2P.20220113.plus15bp.merged.bed # OK
CLINVAR=/home/u035/u035/shared/resources/G2P/DDG2P.20220113.clinvar.20220109.plus15bp.txt # OK
BLACKLIST=/home/u035/u035/shared/resources/blacklist/current_blacklist.txt # OK
TRANS_MAP=/home/u035/u035/shared/resources/trans_map/current_trans_map.txt # OK
REC_SNP=/home/u035/u035/shared/resources/reccurent/current_reccurent.bed # OK, https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7116826/, Extended Data Table 1
### TOOLS ###
SAMTOOLS=/home/u035/u035/shared/software/bcbio/anaconda/bin/samtools
BCFTOOLS=/home/u035/u035/shared/software/bcbio/anaconda/envs/python2/bin/bcftools
BGZIP=/home/u035/u035/shared/software/bcbio/anaconda/envs/python2/bin/bgzip
TABIX=/home/u035/u035/shared/software/bcbio/anaconda/envs/python2/bin/tabix
VT=/home/u035/u035/shared/software/bcbio/anaconda/bin/vt
VASE=/home/u035/u035/shared/software/bcbio/anaconda/bin/vase
GATK4=/home/u035/u035/shared/software/bcbio/anaconda/bin/gatk # points to ../share/gatk4-4.2.1.0-0/gatk
GATK3=/home/u035/u035/shared/software/GenomeAnalysisTK-3.8/GenomeAnalysisTK.jar
PYTHON3=/home/u035/u035/shared/software/bcbio/anaconda/bin/python3 # points to python3.6
PYTHON2=/home/u035/u035/shared/software/bcbio/anaconda/envs/python2/bin/python2.7
VEP="/home/u035/u035/shared/software/bcbio/anaconda/bin/perl /home/u035/u035/shared/software/bcbio/anaconda/bin/vep" # points to ../share/ensembl-vep-100.4-0/vep
REFERENCE_GENOME=/home/u035/u035/shared/software/bcbio/genomes/Hsapiens/hg38/seq/hg38.fa
echo "SOURCE_DIR = ${SOURCE_DIR}" # the general path to the source BAM files (VCF and PED already copied) i.e. /home/u035/u035/shared/results
echo "BATCH_ID = ${BATCH_ID}" # the ID of the batch being processed e.g. 19650_Ansari_Morad
echo "BATCH_NUM = ${BATCH_NUM}" # the numerical part of the BATCH_ID e.g. 19650
echo "PLATE_ID = ${PLATE_ID}" # the PCR plate ID of the batch being currently processed, e.g. 19285
echo "PROJECT_ID = ${PROJECT_ID}" # this the the folder (${BASE}/${PROJECT_ID}) where the downstream analysis will be done
echo "VERSION_N = ${VERSION_N}" # the version of the alignment and genotyping analysis
# change to the LOG folder
cd ${LOG_DIR}
################################
##### for each family ####
################################
FAMILY_ID=`head -n ${SLURM_ARRAY_TASK_ID} ${FAMILY_IDS} | tail -n 1`
########################################################################
### DNU and clean the each family VCF ###
### format: ${PLATE_ID}_${FAMILY_ID}-gatk-haplotype-annotated.vcf.gz ###
########################################################################
echo "Performing DNU and cleaning of the ${PLATE_ID}_${FAMILY_ID}'s VCF file..."
time ${VT} decompose -s ${VCF_DIR}/${PLATE_ID}_${FAMILY_ID}-gatk-haplotype-annotated.vcf.gz -o ${VCF_DIR}/${PLATE_ID}_${FAMILY_ID}.decomp.vcf.gz
time ${VT} normalize ${VCF_DIR}/${PLATE_ID}_${FAMILY_ID}.decomp.vcf.gz -r ${REFERENCE_GENOME} -o ${VCF_DIR}/${PLATE_ID}_${FAMILY_ID}.norm.vcf.gz
time ${VT} uniq ${VCF_DIR}/${PLATE_ID}_${FAMILY_ID}.norm.vcf.gz -o ${VCF_DIR}/${PLATE_ID}_${FAMILY_ID}.DNU.vcf.gz
# remove sites with AC=0
time ${BCFTOOLS} view --min-ac=1 --no-update ${VCF_DIR}/${PLATE_ID}_${FAMILY_ID}.DNU.vcf.gz > ${VCF_DIR}/${PLATE_ID}_${FAMILY_ID}.AC0.vcf
# reset GT to no-call if num_ALT < num_ALT_THERSH or VAF < VAF_THRESH and GT != 0/0
# exlude variants from the blacklist (matching on chr,pos,ref,alt)
time ${PYTHON2} ${SCRIPTS_DIR}/filter_LQ_GT.py ${BLACKLIST} ${VCF_DIR}/${PLATE_ID}_${FAMILY_ID}.AC0.vcf ${VCF_DIR}/${PLATE_ID}_${FAMILY_ID}.clean.vcf
# bgzip and tabix it
time cat ${VCF_DIR}/${PLATE_ID}_${FAMILY_ID}.clean.vcf | ${BGZIP} > ${VCF_DIR}/${PLATE_ID}_${FAMILY_ID}.ready.vcf.gz
time ${TABIX} -p vcf ${VCF_DIR}/${PLATE_ID}_${FAMILY_ID}.ready.vcf.gz
# delete intermediate files
rm ${VCF_DIR}/${PLATE_ID}_${FAMILY_ID}-gatk-haplotype-annotated.vcf.gz
rm ${VCF_DIR}/${PLATE_ID}_${FAMILY_ID}.decomp.vcf.gz*
rm ${VCF_DIR}/${PLATE_ID}_${FAMILY_ID}.norm.vcf.gz*
rm ${VCF_DIR}/${PLATE_ID}_${FAMILY_ID}.DNU.vcf.gz*
rm ${VCF_DIR}/${PLATE_ID}_${FAMILY_ID}.AC0.vcf
# to avoid bgzip pipe broken annoying, but not problematic message - skip the next step, the file will be used by G2P as IN_FILE and will be deleted last
# rm ${VCF_DIR}/${PLATE_ID}_${FAMILY_ID}.clean.vcf
echo ""
echo ""
echo "^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^"
echo "DNU, AC=0, num_ALT & VAF & blacklist cleaning and of the ${PLATE_ID}_${FAMILY_ID}'s VCF file: done"
echo "^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^"
echo ""
echo ""
###########################################################
### run G2P for each family VCF (DD genes) ###
### format: ${PLATE_ID}_${FAMILY_ID}.clean.vcf ###
###########################################################
echo "Performing G2P analysis (DD genes)for FAMILY_ID = ${PLATE_ID}_${FAMILY_ID}..."
echo "Using ${TARGETS}"
IN_FILE=${VCF_DIR}/${PLATE_ID}_${FAMILY_ID}.clean.vcf
G2P_LOG_DIR=${G2P_DIR}/${PLATE_ID}_${FAMILY_ID}_LOG_DIR
mkdir ${G2P_LOG_DIR}
TXT_OUT=${G2P_LOG_DIR}/${PLATE_ID}_${FAMILY_ID}.report.txt
HTML_OUT=${G2P_LOG_DIR}/${PLATE_ID}_${FAMILY_ID}.report.html
VCF_KEYS='gnomADe_r2.1.1_GRCh38|gnomADg_r3.1.1_GRCh38'
time ${VEP} \
-i ${IN_FILE} \
--output_file ${G2P_LOG_DIR}/${PLATE_ID}_${FAMILY_ID}_inter_out.txt \
--force_overwrite \
--assembly GRCh38 \
--fasta ${REFERENCE_GENOME} \
--offline \
--merged \
--use_given_ref \
--cache --cache_version 100 \
--dir_cache /home/u035/u035/shared/software/bcbio/genomes/Hsapiens/hg38/vep \
--individual all \
--transcript_filter "gene_symbol in /home/u035/u035/shared/resources/G2P/genes_in_DDG2P.20220113.txt" \
--dir_plugins /home/u035/u035/shared/software/bcbio/anaconda/share/ensembl-vep-100.4-0 \
--plugin G2P,file='/home/u035/u035/shared/resources/G2P/DDG2P.20220113.csv',af_from_vcf=1,confidence_levels='definitive&strong',af_from_vcf_keys=${VCF_KEYS},log_dir=${G2P_LOG_DIR},txt_report=${TXT_OUT},html_report=${HTML_OUT}
echo ""
echo ""
echo "^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^"
echo "G2P analysis of FAMILY_ID = ${PLATE_ID}_${FAMILY_ID}: done"
echo "^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^"
echo ""
echo ""
############################################################################################################################################################################################
### run coverage for each proband (DD genes) ###
### format: ${SOURCE_DIR}/${BATCH_NUM}_${VERSION_N}/families/????-??-??_${BATCH_NUM}_${VERSION_N}_${PLATE_ID}_${FAMILY_ID}/${INDI_ID}_${FAMILY_ID}/${INDI_ID}_${FAMILY_ID}-ready.bam ###
############################################################################################################################################################################################
#################################
##### for each proband ####
#################################
PROBAND_ID=`head -n ${SLURM_ARRAY_TASK_ID} ${CHILD_IDS} | tail -n 1` # contains only the proband IDs (e.g. 107060)
echo "Performing coverage analysis for PROBAND_ID = ${PROBAND_ID}_${FAMILY_ID} ..."
# make sure we are reading the data from the exact batch & plate ID
BAM_FILE=${SOURCE_DIR}/${BATCH_NUM}_${VERSION_N}/families/????-??-??_${BATCH_NUM}_${VERSION_N}_${PLATE_ID}_${FAMILY_ID}/${PROBAND_ID}_${FAMILY_ID}/${PROBAND_ID}_${FAMILY_ID}-ready.bam
OUT_FILE=${COV_DIR}/${PROBAND_ID}_${FAMILY_ID}.DD15
time java -Xmx8g -jar ${GATK3} -T DepthOfCoverage -R ${REFERENCE_GENOME} -o ${OUT_FILE} -I ${BAM_FILE} -L ${TARGETS} \
--omitDepthOutputAtEachBase \
--minBaseQuality 20 \
--minMappingQuality 20 \
-ct 20 \
-jdk_deflater \
-jdk_inflater \
--allow_potentially_misencoded_quality_scores
echo ""
echo ""
echo "----------------------------------------------------------------------------------------------------"
echo "percentage of DD exons (+/-15bp) covered at least 20x in PROBAND_ID = ${PROBAND_ID}_${FAMILY_ID} ..."
cat ${COV_DIR}/${PROBAND_ID}_${FAMILY_ID}.DD15.sample_summary | awk '{print $7}'
echo "----------------------------------------------------------------------------------------------------"
# now compute the coverage per DD exon (+/-15bp) interval, adding the number of P/LP ClinVar variants (assertion criteria provided) in each interval
time ${PYTHON2} ${SCRIPTS_DIR}/get_cov_output.py ${COV_DIR}/${PROBAND_ID}_${FAMILY_ID}.DD15.sample_interval_summary ${CLINVAR} ${COV_DIR}/${PROBAND_ID}_${FAMILY_ID}.DD15.COV.txt
echo ""
echo ""
echo "^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^"
echo "Coverage analysis of PROBAND_ID = ${PROBAND_ID}_${FAMILY_ID}: done "
echo " Coverage file = ${COV_DIR}/${PROBAND_ID}_${FAMILY_ID}.DD15.COV.txt"
echo "^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^"
echo ""
echo ""
################################################################################################
# check the coverage per each of the reccurent de novo SNPs (padded with 15bp both directions) #
################################################################################################
echo "Performing recurrent coverage analysis for PROBAND_ID = ${PROBAND_ID}_${FAMILY_ID} ..."
# we have identified the name of the proband's BAM file above (BAM_FILE), reuse it
# set the name of the file containing info about the coverage of the recurrent SNPs
REC_OUT_FILE=${COV_DIR}/${PROBAND_ID}_${FAMILY_ID}.REC_SNP_COV.txt
while IFS=$'\t' read -ra var; do
gene="${var[0]}"
chr="${var[1]}"
pos="${var[2]}"
lo=$(expr $pos - 15)
hi=$(expr $pos + 15)
reg="$lo-$hi"
echo "============================================="
echo "$gene : recurrent variant at $chr:$pos"
echo "exploring coverage at $chr:$reg"
echo "---------------------------------------------"
echo "precisely at the position"
${SAMTOOLS} depth -aa -Q 20 -r $chr:$reg ${BAM_FILE} | grep "$pos"
echo "---------------------------------------------"
echo "average in the +/- 15bp region"
${SAMTOOLS} depth -aa -Q 20 -r $chr:$reg ${BAM_FILE} | awk '{sum+=$3} END { print "Average = ",sum/NR}'
echo "---------------------------------------------"
echo "detailed in the +/- 15bp region"
${SAMTOOLS} depth -aa -Q 20 -r $chr:$reg ${BAM_FILE}
done < ${REC_SNP} > ${REC_OUT_FILE}
echo ""
echo ""
echo "^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^"
echo "Coverage analysis of recurring SNPs for PROBAND_ID = ${PROBAND_ID}_${FAMILY_ID}: done "
echo " Coverage file = ${REC_OUT_FILE}"
echo "^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^"
echo ""
echo ""
###################################################################################
### for each proband generate the DECIPHER file ###
### ${VCF_DIR}/${PLATE_ID}_${FAMILY_ID}.ready.vcf.gz - the cleaned family VCF ###
### ${VASE_DIR}/${PLATE_ID}_${FAMILY_ID}.ready.denovo.vcf - the VASE file ###
### ${TRANS_MAP} - the current transcript mapping file ###
###################################################################################
echo "Generating the DECIPHER file for PROBAND_ID = ${PROBAND_ID}_${FAMILY_ID} ..."
# first, split the family VCF to individual VCFs
# -c1: minimum allele count (INFO/AC) of sites to be printed
# split multi-allelic sites (by -m -any)
# left-alignment and normalization (by adding the -f)
file=${VCF_DIR}/${PLATE_ID}_${FAMILY_ID}.ready.vcf.gz
echo "splitting $file"
for indi in `${BCFTOOLS} query -l $file`; do
${BCFTOOLS} view -c1 -Oz -s $indi -o ${file/.vcf*/.$indi.rough.vcf.gz} $file
${BCFTOOLS} norm -f ${REFERENCE_GENOME} -m -any -Oz -o ${file/.vcf*/.$indi.vcf.gz} ${file/.vcf*/.$indi.rough.vcf.gz}
rm ${file/.vcf*/.$indi.rough.vcf.gz}
done
################# VASE file - already split, left-aligned and normalized
# create the names of the needed files
PED_FILE=${PED_DIR}/${BATCH_ID}_${PLATE_ID}_${FAMILY_ID}.ped
DEC_MAP=${WORK_DIR}/solo_DECIPHER_INTERNAL_IDs.txt
IN_G2P_FILE=${G2P_DIR}/${PLATE_ID}_${FAMILY_ID}_LOG_DIR/${PLATE_ID}_${FAMILY_ID}.report.txt
FAM_IGV_DIR=${IGV_DIR}/${PLATE_ID}_${FAMILY_ID}
FAM_BAM_DIR=${SOURCE_DIR}/${BATCH_NUM}_${VERSION_N}/families/????-??-??_${BATCH_NUM}_${VERSION_N}_${PLATE_ID}_${FAMILY_ID}
## call the python scrpit
time ${PYTHON2} ${SCRIPTS_DIR}/generate_DEC_IGV_solo_scripts.py \
${DEC_MAP} \
${TRANS_MAP} \
${PED_FILE} \
${IN_G2P_FILE} \
${FAM_IGV_DIR} \
${VCF_DIR} \
${PLATE_ID} \
${FAMILY_ID} \
${DEC_DIR} \
${FAM_BAM_DIR}
## using the DECIPHER bulk upload file v9 --> generate the DECIPHER bulk upload file v10
echo "...Generating v10 Decipher bulk upload file for proband = ${PROBAND_ID}, family_id = ${FAMILY_ID} ..."
time ${PYTHON3} ${SCRIPTS_DIR}/convert_DEC_to_v10.py ${DEC_DIR} ${PROBAND_ID}_${FAMILY_ID}
echo ""
echo ""
echo "^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^"
echo "DECIPHER analysis of PROBAND_ID = ${PROBAND_ID}_${FAMILY_ID}: done"
echo "^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^"
echo ""
echo ""
rm ${VCF_DIR}/${PLATE_ID}_${FAMILY_ID}.clean.vcf
##############################################################################################
### for each variant in the DECIPHER upload file ###
### generate a IGV snapshot based on the realigned BAM used by GATK for calling variants ###
### first, generate BAMOUTs for each variant (to be stored in the BAMOUT folder) ###
### then, generate a batch script for IGV to produce the snapshots based on the BAMOUTs ###
##############################################################################################
# we have so far
# FAMILY_ID=`head -n ${SLURM_ARRAY_TASK_ID} ${FAMILY_IDS} | tail -n 1`
# PROBAND_ID=`head -n ${SLURM_ARRAY_TASK_ID} ${CHILD_IDS} | tail -n 1`
echo "...Generating BAMOUT files for the ${FAMILY_ID} family, proband = ${PROBAND_ID} ..."
# identify proband ID from the VCF file
kid_id=''
file=${VCF_DIR}/${PLATE_ID}_${FAMILY_ID}.ready.vcf.gz
for indi in `${BCFTOOLS} query -l $file`; do
echo "indi = $indi"
if [ "${indi}" = "${PROBAND_ID}_${FAMILY_ID}" ]
then
kid_id=${indi}
fi
done
echo "...kid_id = ${kid_id} "
# gather the proband BAM file
kid_bam=${SOURCE_DIR}/${BATCH_NUM}_${VERSION_N}/families/????-??-??_${BATCH_NUM}_${VERSION_N}_${PLATE_ID}_${FAMILY_ID}/${kid_id}/${kid_id}-ready.bam
echo "...kid_bam = ${kid_bam}..."
# gather the variants in the DECIPHER file for which to generate bamouts
# chr is the second column - need to add the 'chr' prefix
# pos is the third column
# the first line is a header line, starting with 'Internal reference number or ID'
# file called: ${DEC_DIR}/<proband_id>_<fam_id>_DEC_FLT.csv
# and for each run GATK to generate the bamout files
# to be stored in ${BAMOUT_DIR}/${FAMILY_ID}
mkdir ${BAMOUT_DIR}/${FAMILY_ID}
var_file=${DEC_DIR}/${kid_id}_DEC_FLT.csv
echo "... reading ${var_file} to generate the bamouts..."
grep -v '^Internal' ${var_file} |
while IFS= read -r line
do
echo "$line"
IFS=, read -ra ary <<<"$line"
# for key in "${!ary[@]}"; do echo "$key ${ary[$key]}"; done
chr=${ary[1]}
pos=${ary[2]}
ref=${ary[4]}
alt=${ary[5]}
echo " --> chr = $chr, pos = $pos, ref = ${ref}, alt = ${alt}"
# generate the bamout file
echo "...doing the bamout"
echo " time ${GATK4} HaplotypeCaller --reference ${REFERENCE_GENOME} --input ${kid_bam} -L chr${chr}:${pos} --interval-padding 500 \"
echo " --active-probability-threshold 0.000 -ploidy 2 \"
echo " --output ${BAMOUT_DIR}/${FAMILY_ID}/${FAMILY_ID}_chr${chr}_${pos}.bamout.vcf -bamout ${BAMOUT_DIR}/${FAMILY_ID}/${FAMILY_ID}_chr${chr}_${pos}.bamout.bam"
time ${GATK4} HaplotypeCaller --reference ${REFERENCE_GENOME} --input ${kid_bam} -L chr${chr}:${pos} \
--interval-padding 500 --active-probability-threshold 0.000 -ploidy 2 \
--output ${BAMOUT_DIR}/${FAMILY_ID}/${FAMILY_ID}_chr${chr}_${pos}.bamout.vcf \
-bamout ${BAMOUT_DIR}/${FAMILY_ID}/${FAMILY_ID}_chr${chr}_${pos}.bamout.bam
done
#################################################################
# write the IGV batch file for this family based on the bamouts #
# to be stored as /home/u035/u035/shared/analysis/work/${PROJECT_ID}/DECIPHER/IGV/bamout_${PROBAND_ID}_${FAMILY_ID}.snapshot.txt #
#################################################################
snap_file=/home/u035/u035/shared/analysis/work/${PROJECT_ID}/DECIPHER/IGV/bamout_${PROBAND_ID}_${FAMILY_ID}.solo.snapshot.txt
# check if previous version exist, if so - delete it
if [ -f "${snap_file}" ]; then
echo "previous version of ${snap_file} exist --> deleted"
rm ${snap_file}
fi
# write the header for the IGV batch file
echo "new" >> ${snap_file}
echo "genome hg38" >> ${snap_file}
echo "snapshotDirectory \"/home/u035/u035/shared/analysis/work/${PROJECT_ID}/DECIPHER/IGV/${PLATE_ID}_${FAMILY_ID}\"" >> ${snap_file}
echo "" >> ${snap_file}
# now, go again over the variants in the DECIPHER file and generate one snapshot file for all the variants
var_file=${DEC_DIR}/${kid_id}_DEC_FLT.csv
echo "... reading ${var_file} to generate the IGV batch file using the bamouts..."
grep -v '^Internal' ${var_file} |
while IFS= read -r line
do
IFS=, read -ra ary <<<"$line"
chr=${ary[1]}
pos=${ary[2]}
ref=${ary[4]}
alt=${ary[5]}
left=$((${pos}-25))
right=$((${pos}+25))
echo "new" >> ${snap_file}
echo "load ${BAMOUT_DIR}/${FAMILY_ID}/${FAMILY_ID}_chr${chr}_${pos}.bamout.bam" >> ${snap_file}
echo "preference SAM.SHADE_BASE_QUALITY true" >> ${snap_file}
echo "goto chr${chr}:${left}-${right}" >> ${snap_file}
echo "group SAMPLE" >> ${snap_file}
echo "sort base" >> ${snap_file}
echo "squish" >> ${snap_file}
echo "snapshot bamout_${PROBAND_ID}_${FAMILY_ID}_chr${chr}_${pos}_${ref}_${alt}.png" >> ${snap_file}
echo "" >> ${snap_file}
echo "" >> ${snap_file}
done
echo "Generating of the IGV batch files based on bamouts - done!"
echo "snap_file = ${snap_file}"
echo "+++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++"
echo "+++ Variant prioritization of family ${FAMILY_ID} completed +++"
echo "+++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++"