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lumpyexpress
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executable file
·559 lines (493 loc) · 16.9 KB
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#!/bin/bash -e
############################################################
# Program: lumpyexpress
# Author: Colby Chiang (cc2qe@virginia.edu)
############################################################
set -eo pipefail
# helper functions to differentiate bam from cram input files. Courtesy idas (https://github.com/indraniel)
# Check if a file is a cram or bam based on the binary header
function is_cram () {
local file_path=$1
header=$($HEXDUMP -n 4 -e '"%c"' ${file_path})
if [ ${header} == 'CRAM' ]; then
result=0
else
result=1
fi
return ${result}
}
function is_bam () {
local file_path=$1
local result=1
is_gzipped=$($HEXDUMP -n 2 -e '"%x"' ${file_path})
if [ ${is_gzipped} = '8b1f' ]; then
is_bam=$(zcat ${file_path} | $HEXDUMP -n 3 -e '"%c"')
if [ ${is_bam} = 'BAM' ]; then
result=0
fi
fi
return ${result}
}
# ensure that we have a valid CRAM or BAM file
function check_valid_cram_or_bam () {
local input_path=$1
if is_cram ${input_path} || is_bam ${input_path} ; then
echo -e "${input_path} is a valid CRAM/BAM file"
else
echo -e "${input_path}: is NOT a valid CRAM/BAM file!"
exit 1
fi
}
# source the paths to the binaries used in the script
source_binaries() {
if [[ -e $1 ]]
then
echo "Sourcing executables from $1 ..."
if [[ $1 == /* ]]
then
source $1
else
source ./$1
fi
else
echo "Config file $1 not found. Attempting to auto-source executables"
# general
LUMPY_HOME=$( dirname `which lumpyexpress` )
LUMPY=`which lumpy || true`
SAMBLASTER=`which samblaster || true`
SAMBAMBA=`which sambamba || true`
SAMTOOLS=`which samtools || true`
# python 2.7 or newer, must have pysam, numpy installed
PYTHON=`which python || true`
# For testing if the full BAM is actually a CRAM!
HEXDUMP=`which hexdump || true`
# python scripts
PAIREND_DISTRO=$LUMPY_HOME/scripts/pairend_distro.py
BAMGROUPREADS=$LUMPY_HOME/scripts/bamkit/bamgroupreads.py
BAMFILTERRG=$LUMPY_HOME/scripts/bamkit/bamfilterrg.py
BAMLIBS=$LUMPY_HOME/scripts/bamkit/bamlibs.py
fi
# sambamba has a bug where it can't parse CRAM (at least on our system)
# see https://github.com/lomereiter/sambamba/issues/280
unset SAMBAMBA
}
# ensure that the require python modules are installed before
# beginning analysis
check_python_modules() {
PYTHON_TEST=$1
echo -e "\nChecking for required python modules ($PYTHON_TEST)..."
$PYTHON_TEST -c "import imp; imp.find_module('pysam')"
$PYTHON_TEST -c "import imp; imp.find_module('numpy')"
}
## usage
usage() {
echo "
usage: lumpyexpress [options]
options:
-B FILE full BAM or CRAM file(s) (comma separated) (required)
-S FILE split reads BAM file(s) (comma separated)
-D FILE discordant reads BAM files(s) (comma separated)
-R FILE indexed reference genome fasta file (recommended for CRAMs)
-d FILE bedpe file of depths (comma separated and prefixed by sample:)
e.g sample_x:/path/to/sample_x.bedpe,sample_y:/path/to/sample_y.bedpe
-o FILE output file [fullBam.bam.vcf]
-x FILE BED file to exclude
-P output probability curves for each variant
-m INT minimum sample weight for a call [4]
-r FLOAT trim threshold [0]
-T DIR temp directory [./output_prefix.XXXXXXXXXXXX]
-k keep temporary files
-K FILE path to lumpyexpress.config file
(default: same directory as lumpyexpress)
-v verbose
-h show this message
"
}
# set defaults
LUMPY_DIR=`dirname $0`
CONFIG="$LUMPY_DIR/lumpyexpress.config"
THREADS=1
ANNOTATE=0
MIN_SAMPLE_WEIGHT=4
TRIM_THRES=0
EXCLUDE_BED=
TEMP_DIR=""
GENOTYPE=0
READDEPTH=0
VERBOSE=0
KEEP=0
OUTPUT=""
MAX_SPLIT_COUNT=2
MIN_NON_OVERLAP=20
PROB_CURVE=""
REF=""
REF_ARG=""
SPL_BAM_STRING=""
DISC_BAM_STRING=""
DEPTH_BED_STRING=""
while getopts ":hB:S:D:R:d:o:m:r:x:T:t:PAdgkvK:" OPTION
do
case "${OPTION}" in
h)
usage
exit 0
;;
B)
FULL_BAM_STRING="$OPTARG"
;;
S)
SPL_BAM_STRING="$OPTARG"
;;
D)
DISC_BAM_STRING="$OPTARG"
;;
R)
REF="$OPTARG"
REF_ARG="-T $OPTARG"
;;
d)
DEPTH_BED_STRING="$OPTARG"
;;
o)
OUTPUT="$OPTARG"
;;
m)
MIN_SAMPLE_WEIGHT="$OPTARG"
;;
r)
TRIM_THRES="$OPTARG"
;;
x)
EXCLUDE_BED="$OPTARG"
EXCLUDE_BED_FMT="-x $EXCLUDE_BED"
;;
T)
TEMP_DIR="$OPTARG"
;;
t)
THREADS="$OPTARG"
;;
P)
PROB_CURVE="-P"
;;
A)
ANNOTATE=1
;;
d)
READDEPTH=1
;;
g)
GENOTYPE=1
;;
v)
VERBOSE=1
;;
k)
KEEP=1
;;
K)
CONFIG="$OPTARG"
;;
esac
done
# parse the BAM strings
FULL_BAM_LIST=($(echo $FULL_BAM_STRING | tr "," " "))
SPL_BAM_LIST=($(echo $SPL_BAM_STRING | tr "," " "))
DISC_BAM_LIST=($(echo $DISC_BAM_STRING | tr "," " "))
DEPTH_BED_LIST=($(echo $DEPTH_BED_STRING | tr "," " "))
OPTIND=0
# Check the for the relevant binaries
source_binaries $CONFIG
if [[ -z "$LUMPY" ]]
then
usage
echo -e "Error: lumpy executable not found. Please set path in $LUMPY_DIR/lumpyexpress.config file\n"
exit 1
elif [[ -z "$PAIREND_DISTRO" ]]
then
usage
echo -e "Error: pairend_distro.py executable not found. Please set path in $LUMPY_DIR/lumpyexpress.config file\n"
exit 1
elif [[ -z "$BAMFILTERRG" ]]
then
usage
echo -e "Error: bamfilterrg.py executable not found. Please set path in $LUMPY_DIR/lumpyexpress.config file\n"
exit 1
fi
# $SAMT will be either sambamba or samtools, depending on which is available
if [[ ! -z "$SAMBAMBA" ]]
then
SAMT="$SAMBAMBA"
CRAM_SAMT_ARG="-C"
SAMT_STREAM="$SAMBAMBA view -f bam -l 0"
SAMTOBAM="$SAMBAMBA view -S -f bam -l 0"
SAMSORT="$SAMBAMBA sort -m 1G --tmpdir "
elif [[ ! -z "$SAMTOOLS" ]]
then
SAMT="$SAMTOOLS"
CRAM_SAMT_ARG=""
SAMT_STREAM="$SAMTOOLS view -u"
SAMTOBAM="$SAMTOOLS view -S -u"
SAMSORT="$SAMTOOLS sort -m 1G -T "
else
usage
echo -e "Error: neither samtools nor sambamba were found. Please set path of one of these in $LUMPY_DIR/lumpyexpress.config file\n"
exit 1
fi
# check for required python modules (pysam, numpy)
check_python_modules $PYTHON
# Check that the required files exist
if [[ ${#FULL_BAM_LIST[@]} -eq 0 ]]
then
usage
echo -e "Error: -B is required\n"
exit 1
fi
set +o nounset
for TEST_BAM in ${FULL_BAM_LIST[@]} ${SPL_BAM_LIST[@]} ${DISC_BAM_LIST[@]}
do
if [[ ! -f $TEST_BAM ]]
then
usage
echo -e "Error: file $TEST_BAM not found.\n"
exit 1
fi
done
for TEST_BED in ${DEPTH_BED_LIST[@]}
do
if [[ -z $(echo "$TEST_BED" | grep ":") ]]
then
usage
echo -e "Error: must specify depths as sample_id:bedpe"
exit 1;
fi
bpath=$(echo "$TEST_BED" | perl -pe 's/^.+://')
if [[ ! -f "$bpath" ]]; then
usage
echo -e "Error: depth bed does not exist: $bpath"
exit 1
fi
done
set -o nounset
# default OUTPUT if not provided
if test -z "$OUTPUT"
then
OUTPUT=`basename "${FULL_BAM_LIST[0]}"`.vcf
fi
OUTBASE=`basename "$OUTPUT"`
# make temporary directory
if [[ $VERBOSE -eq 1 ]]
then
echo "
create temporary directory"
fi
if [[ -z $TEMP_DIR ]]
then
TEMP_DIR=`mktemp -d ${OUTBASE}.XXXXXXXXXXXX`
else
mkdir -p $TEMP_DIR
fi
cleanup () {
rm -rf $TEMP_DIR
}
trap cleanup EXIT
# If splitter and discordant BAMs not provided, generate them
# (LUMPY express)
set +o nounset
if [[ -z "${SPL_BAM_LIST}${DISC_BAM_LIST}" ]]
then
# initialize split and discordant bam lists
SPL_BAM_LIST=()
DISC_BAM_LIST=()
# create temp files and pipes
mkdir -p $TEMP_DIR/spl $TEMP_DIR/disc
if [[ ! -e $TEMP_DIR/spl_pipe ]]
then
mkfifo $TEMP_DIR/spl_pipe
fi
if [[ ! -e $TEMP_DIR/disc_pipe ]]
then
mkfifo $TEMP_DIR/disc_pipe
fi
# generate histo files and construct the strings for LUMPY
for i in $( seq 0 $(( ${#FULL_BAM_LIST[@]}-1 )) )
do
FULL_BAM=${FULL_BAM_LIST[$i]}
EXT_OPT=""
if is_cram $FULL_BAM; then
EXT_OPT="$CRAM_SAMT_ARG $REF_ARG"
fi
# calc readlength if not provided
set +o pipefail
READ_LENGTH=`$SAMT view $EXT_OPT $FULL_BAM | head -n 10000 | gawk 'BEGIN { MAX_LEN=0 } { LEN=length($10); if (LEN>MAX_LEN) MAX_LEN=LEN } END { print MAX_LEN }'`
set -o pipefail
# parse the libraries in the BAM header to extract readgroups from the same library
LIB_RG_LIST=(`$PYTHON $BAMLIBS $FULL_BAM`)
# process each library's splitters and discordants
for j in $( seq 0 $(( ${#LIB_RG_LIST[@]}-1 )) )
do
if [[ "$VERBOSE" -eq 1 ]]
then
echo -e "
$PYTHON $BAMGROUPREADS --fix_flags -i $FULL_BAM -r ${LIB_RG_LIST[$j]} \\
| $SAMBLASTER --acceptDupMarks --excludeDups --addMateTags --maxSplitCount $MAX_SPLIT_COUNT --minNonOverlap $MIN_NON_OVERLAP \\
--splitterFile $TEMP_DIR/spl_pipe --discordantFile $TEMP_DIR/disc_pipe \\
| $SAMT view -S /dev/stdin \\
| gawk '{ if (NR<=1000000) print > \"/dev/stdout\" ; else print > \"/dev/null\" }' \\
| $PYTHON $PAIREND_DISTRO -r $READ_LENGTH -X 4 -N 1000000 -o ${TEMP_DIR}/$OUTBASE.sample$(($i+1)).lib$(($j+1)).x4.histo \\
> ${TEMP_DIR}/$OUTBASE.sample$(($i+1)).lib$(($j+1)).insert.stats
$SAMTOBAM $TEMP_DIR/spl_pipe \\
| $SAMSORT $TEMP_DIR/spl -o $TEMP_DIR/$OUTBASE.sample$(($i+1)).lib$(($j+1)).splitters.bam /dev/stdin
$SAMTOBAM $TEMP_DIR/disc_pipe \\
| $SAMSORT $TEMP_DIR/disc -o $TEMP_DIR/$OUTBASE.sample$(($i+1)).lib$(($j+1)).discordants.bam /dev/stdin"
fi
$PYTHON $BAMGROUPREADS --fix_flags -i $FULL_BAM -r ${LIB_RG_LIST[$j]} \
| $SAMBLASTER --acceptDupMarks --excludeDups --addMateTags --maxSplitCount $MAX_SPLIT_COUNT --minNonOverlap $MIN_NON_OVERLAP \
--splitterFile $TEMP_DIR/spl_pipe --discordantFile $TEMP_DIR/disc_pipe \
| $SAMT view -S /dev/stdin \
| gawk '{ if (NR<=1000000) print > "/dev/stdout" ; else print > "/dev/null" }' \
| $PYTHON $PAIREND_DISTRO -r $READ_LENGTH -X 4 -N 1000000 -o ${TEMP_DIR}/$OUTBASE.sample$(($i+1)).lib$(($j+1)).x4.histo \
> ${TEMP_DIR}/$OUTBASE.sample$(($i+1)).lib$(($j+1)).insert.stats &
$SAMTOBAM $TEMP_DIR/spl_pipe \
| $SAMSORT $TEMP_DIR/spl -o $TEMP_DIR/$OUTBASE.sample$(($i+1)).lib$(($j+1)).splitters.bam /dev/stdin &
$SAMTOBAM $TEMP_DIR/disc_pipe \
| $SAMSORT $TEMP_DIR/disc -o $TEMP_DIR/$OUTBASE.sample$(($i+1)).lib$(($j+1)).discordants.bam /dev/stdin
wait
# generate discordant pair string for LUMPY
DISC_BAM=$TEMP_DIR/$OUTBASE.sample$(($i+1)).discordants.bam
DISC_SAMPLE=`$SAMT view $EXT_OPT -H $FULL_BAM | grep -m 1 "^@RG" | gawk -v i=$i '{ for (j=1;j<=NF;++j) {if ($j~"^SM:") { gsub("^SM:","",$j); print $j } } }'`
RG_STRING=`echo "${LIB_RG_LIST[$j]}" | sed 's/,/,read_group:/g' | sed 's/^/read_group:/g'`
MEAN=`cat ${TEMP_DIR}/$OUTBASE.sample$(($i+1)).lib$(($j+1)).insert.stats | tr '\t' '\n' | grep "^mean" | sed 's/mean\://g'`
STDEV=`cat ${TEMP_DIR}/$OUTBASE.sample$(($i+1)).lib$(($j+1)).insert.stats | tr '\t' '\n' | grep "^stdev" | sed 's/stdev\://g'`
LUMPY_DISC_STRING="$LUMPY_DISC_STRING -pe bam_file:${DISC_BAM},histo_file:${TEMP_DIR}/$OUTBASE.sample$(($i+1)).lib$(($j+1)).x4.histo,mean:${MEAN},stdev:${STDEV},read_length:${READ_LENGTH},min_non_overlap:${READ_LENGTH},discordant_z:5,back_distance:10,weight:1,id:${DISC_SAMPLE},min_mapping_threshold:20,${RG_STRING}"
# generate split-read string for LUMPY
SPL_BAM=$TEMP_DIR/$OUTBASE.sample$(($i+1)).splitters.bam
SPL_SAMPLE=`$SAMT view $EXT_OPT -H $FULL_BAM | grep -m 1 "^@RG" | gawk -v i=$i '{ for (j=1;j<=NF;++j) {if ($j~"^SM:") { gsub("^SM:","",$j); print $j } } }'`
LUMPY_SPL_STRING="$LUMPY_SPL_STRING -sr bam_file:${SPL_BAM},back_distance:10,min_mapping_threshold:20,weight:1,id:${SPL_SAMPLE},min_clip:20,${RG_STRING}"
done
# merge the splitters and discordants files
if [[ ${#LIB_RG_LIST[@]} -gt 1 ]]
then
MERGE_DISCORDANTS=""
MERGE_SPLITTERS=""
for j in $( seq 0 $(( ${#LIB_RG_LIST[@]}-1 )) )
do
MERGE_DISCORDANTS="$MERGE_DISCORDANTS $TEMP_DIR/$OUTBASE.sample$(($i+1)).lib$(($j+1)).discordants.bam"
MERGE_SPLITTERS="$MERGE_SPLITTERS ${TEMP_DIR}/$OUTBASE.sample$(($i+1)).lib$(($j+1)).splitters.bam"
done
if [[ $VERBOSE -eq 1 ]]
then
echo "
$SAMT merge ${TEMP_DIR}/$OUTBASE.sample$(($i+1)).discordants.bam $MERGE_DISCORDANTS
$SAMT merge ${TEMP_DIR}/$OUTBASE.sample$(($i+1)).splitters.bam $MERGE_SPLITTERS
rm $MERGE_DISCORDANTS $MERGE_SPLITTERS"
fi
$SAMT merge ${TEMP_DIR}/$OUTBASE.sample$(($i+1)).discordants.bam $MERGE_DISCORDANTS
$SAMT merge ${TEMP_DIR}/$OUTBASE.sample$(($i+1)).splitters.bam $MERGE_SPLITTERS
rm $MERGE_DISCORDANTS $MERGE_SPLITTERS
else
mv ${TEMP_DIR}/$OUTBASE.sample$(($i+1)).lib$(($j+1)).discordants.bam ${TEMP_DIR}/$OUTBASE.sample$(($i+1)).discordants.bam
mv ${TEMP_DIR}/$OUTBASE.sample$(($i+1)).lib$(($j+1)).splitters.bam ${TEMP_DIR}/$OUTBASE.sample$(($i+1)).splitters.bam
fi
# update the splitters and discordant BAM lists
SPL_BAM_LIST+=(${TEMP_DIR}/$OUTBASE.sample$(($i+1)).splitters.bam)
DISC_BAM_LIST+=(${TEMP_DIR}/$OUTBASE.sample$(($i+1)).discordants.bam)
done
# else (user provided a splitter and discordants file)
else
# parse the libraries in the BAM header to extract readgroups from the same library
for i in $( seq 0 $(( ${#FULL_BAM_LIST[@]}-1 )) )
do
FULL_BAM=${FULL_BAM_LIST[$i]}
EXT_OPT=""
if is_cram $FULL_BAM; then
EXT_OPT="$CRAM_SAMT_ARG $REF_ARG"
fi
DISC_BAM=${DISC_BAM_LIST[$i]}
SPL_BAM=${SPL_BAM_LIST[$i]}
# LIB_RG_LIST contains an element for each library in the BAM file.
# These elements are comma delimited strings for the readgroups for each library.
LIB_RG_LIST=(`$PYTHON $BAMLIBS ${FULL_BAM_LIST[$i]}`)
if [[ ${#LIB_RG_LIST[@]} -eq 0 ]]
then
echo "Warning: BAM file lacks read groups, paired-end analysis may fail"
fi
# generate the histo, stats, and config files
echo "Calculating insert distributions... "
for j in $( seq 0 $(( ${#LIB_RG_LIST[@]}-1 )) )
do
# calculate read length if not provided
set +o pipefail
LIB_READ_LENGTH_LIST+=(`$SAMT view $EXT_OPT ${FULL_BAM_LIST[$i]} | head -n 10000 | gawk 'BEGIN { MAX_LEN=0 } { LEN=length($10); if (LEN>MAX_LEN) MAX_LEN=LEN } END { print MAX_LEN }'`)
echo "Library read groups: ${LIB_RG_LIST[$j]}"
echo "Library read length: ${LIB_READ_LENGTH_LIST[$j]}"
$SAMT_STREAM $EXT_OPT ${FULL_BAM_LIST[$i]} \
| $PYTHON $BAMFILTERRG -n 10000000 --readgroup ${LIB_RG_LIST[$j]} \
| grep -v '^@' \
| tail -n 1000000 \
| $PYTHON $PAIREND_DISTRO -r ${LIB_READ_LENGTH_LIST[$j]} -X 4 -N 1000000 -o ${TEMP_DIR}/$OUTBASE.sample$(($i+1)).lib$(($j+1)).x4.histo \
> ${TEMP_DIR}/$OUTBASE.sample$(($i+1)).lib$(($j+1)).insert.stats
set -o pipefail
done
echo "done"
# construct LUMPY_SPL_STRING
SPL_SAMPLE=`$SAMT view -H $SPL_BAM | grep -m 1 "^@RG" | gawk -v i=$i '{ for (j=1;j<=NF;++j) {if ($j~"^SM:") { gsub("^SM:","",$j); print $j } } }'`
LUMPY_SPL_STRING="$LUMPY_SPL_STRING -sr bam_file:${SPL_BAM},back_distance:10,min_mapping_threshold:20,weight:1,id:${SPL_SAMPLE},min_clip:20"
# construct LUMPY_DISC_STRING
for j in $( seq 0 $(( ${#LIB_RG_LIST[@]}-1 )) )
do
echo $(( ${#FULL_BAM_LIST[@]}-1 ))
DISC_BAM=${DISC_BAM_LIST[$i]}
DISC_SAMPLE=`$SAMT view -H $DISC_BAM | grep -m 1 "^@RG" | gawk -v i=$i '{ for (j=1;j<=NF;++j) {if ($j~"^SM:") { gsub("^SM:","",$j); print $j } } }'`
MEAN=`cat ${TEMP_DIR}/$OUTBASE.sample$(($i+1)).lib$(($j+1)).insert.stats | tr '\t' '\n' | grep "^mean" | sed 's/mean\://g'`
STDEV=`cat ${TEMP_DIR}/$OUTBASE.sample$(($i+1)).lib$(($j+1)).insert.stats | tr '\t' '\n' | grep "^stdev" | sed 's/stdev\://g'`
RG_STRING=`echo "${LIB_RG_LIST[$j]}" | sed 's/,/,read_group:/g' | sed 's/^/read_group:/g'`
LUMPY_DISC_STRING="$LUMPY_DISC_STRING -pe bam_file:${DISC_BAM},histo_file:${TEMP_DIR}/$OUTBASE.sample$(($i+1)).lib$(($j+1)).x4.histo,mean:${MEAN},stdev:${STDEV},read_length:${LIB_READ_LENGTH_LIST[$j]},min_non_overlap:${LIB_READ_LENGTH_LIST[$j]},discordant_z:5,back_distance:10,weight:1,id:${DISC_SAMPLE},min_mapping_threshold:20,${RG_STRING}"
done
done
fi
LUMPY_DEPTH_STRING=""
if [[ ! -z "$DEPTH_BED_LIST" ]]; then
# -bedpe bedpe_file:<bedpe file>,id:<sample name>,weight:<sample weight>
set -o nounset
for j in $( seq 0 $(( ${#DEPTH_BED_LIST[@]}-1 )) )
do
rec=${DEPTH_BED_LIST[$j]}
f=$(echo $rec | perl -pe 's/^.+://')
sample=$(echo $rec | perl -pe 's/:.+$//')
# give weight of 4 since these have been called before.
LUMPY_DEPTH_STRING="$LUMPY_DEPTH_STRING -bedpe bedpe_file:$f,id:$sample,weight:4"
done
set +o nounset
fi
echo "Running LUMPY... "
if [[ "$VERBOSE" -eq 1 ]]
then
echo "
$LUMPY ${PROB_CURVE} \\
-t ${TEMP_DIR}/${OUTBASE} \\
-msw $MIN_SAMPLE_WEIGHT \\
-tt $TRIM_THRES \\
$LUMPY_DEPTH_STRING \\
$EXCLUDE_BED_FMT \\
$LUMPY_DISC_STRING \\
$LUMPY_SPL_STRING \\
> $OUTPUT"
fi
# call lumpy
$LUMPY $PROB_CURVE -t ${TEMP_DIR}/${OUTBASE} -msw $MIN_SAMPLE_WEIGHT -tt $TRIM_THRES \
$LUMPY_DEPTH_STRING \
$EXCLUDE_BED_FMT \
$LUMPY_DISC_STRING \
$EXCLUDE_BED_FMT \
$LUMPY_SPL_STRING \
> $OUTPUT
# clean up
if [[ "$KEEP" -eq 0 ]]
then
rm -r ${TEMP_DIR}
fi
echo "LUMPY Express done"
# exit cleanly
exit 0
## END SCRIPT