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Config
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#!/bin/bash
# More complete information on how to fill out
# this Config file can be found at:
# https://github.com/MorrellLAB/sequence_handling/wiki/Configuration
###################################################
########## Shared across handlers ##########
###################################################
# Where are we storing the output files?
# Final directory is ${OUT_DIR}/Name_of_Handler
OUT_DIR=/home/morrellp/large/soybean_toy-all-libs
# Name this project
PROJECT=soybean_toy_dataset_all_libs
# What email should we use for job notifications?
# What encoding is used for quality values?
# Look at the FastQC files to determine the sequence encoding.
# Choose from: 'sanger', 'illumina', 'solexa', or 'phred'
# Illumina 1.8+ and Oxford Nanopore use 'sanger' encoding.
QUAL_ENCODING=sanger
# Sequencing platform technology
# What platform were the reads produced?
# Valid options are:
# CAPILLARY, LS454, ILLUMINA,
# SOLID, HELICOS, IONTORRENT,
# ONT, and PACBIO
SEQ_PLATFORM=ILLUMINA
# What reference genome are we using?
# Include the full physical file path.
REF_GEN=/home/morrellp/large/soybean_toy/genome/Gmax_275_v2.0.fa
# Is this organism barley?
# Choose from: "true" or "false"
BARLEY=false
# Are you running the analysis on the Minnesota Supercomputing Institute (MSI)?
# Choose from: "true" or "false"
MSI=true
# Are you submitting the job with qsub from PBS (Portable Batch System)
# Choose from: "true" or "false"
USE_PBS=false
# Are you submitting the job with sbatch from Slurm?
# Choose from: "true" or "false"
USE_SLURM=true
# Do the quality scores need to be adjusted for GATK? Default: false
# Change to true if you get errors from GATK like:
# "<Sample> appears to be using the wrong encoding for quality scores: we encountered an extremely high quality score"
FIX_QUALITY_SCORES=false
# Where should Picard/GATK store temporary files?
# If you've encountered issues with running out of temp space
# with picard, you can optionally specify a temp directory
# Otherwise, leave blank
TMP=/home/morrellp/large/soybean_toy-all-libs/tmp
############################################
########## GBS_Demultiplex ##########
############################################
# IMPORTANT: GBS_Demultiplex is not yet functional and is still under development!
# Which queue are we submitting the job to?
# Note: For USE_PBS, we can only specify one queue
# For USE_SLURM, we can specify multiple queues that are comma separated, no spaces (e.g., "small,ram256g,ram1t")
GD_QUEUE="small"
# USE_SLURM: What are our sbatch settings?
# Below are the recommended settings
GD_SBATCH="--nodes=1 --ntasks-per-node=4 --mem=1gb --tmp=1gb -t 06:00:00 --mail-type=ALL --mail-user=${EMAIL} -p ${GD_QUEUE}"
# USE_PBS: What are our QS
GD_QSUB="mem=1gb,nodes=1:ppn=4,walltime=6:00:00"
GBS_SAMPLES=
# A key file with the following columns:
# Flowcell Lane Barcode Sample PlateName Row Column
KEY_FILE=
# Are the barcodes at the beginning or the end of the sequence?
# Choose from: 'beginning' or 'end'
LINE_ENDING=
# How many mismatches are allowed?
MISMATCH_TOL=1
# Do we allow partial overlap?
PARTIAL=0
# What extension or naming convention do the samples have?
FILE_TYPE=
############################################
########## Quality_Assessment ##########
############################################
# Which queue are we submitting the job to?
# Note: For USE_PBS, we can only specify one queue
# For USE_SLURM, we can specify multiple queues that are comma separated, no spaces (e.g., "small,ram256g,ram1t")
QA_QUEUE="small"
# USE_SLURM: What are our sbatch settings?
# Below are the recommended settings
QA_SBATCH="--nodes=1 --ntasks-per-node=4 --mem=1gb --tmp=1gb -t 06:00:00 --mail-type=ALL --mail-user=${EMAIL} -p ${QA_QUEUE}"
# USE_PBS: What are our QSub settings for Quality_Assessment?
# Below are the recommended settings
QA_QSUB="mem=1gb,nodes=1:ppn=4,walltime=6:00:00"
# Provide a list of FastQ, SAM, or BAM files to be used
# Include the full file path.
QA_SAMPLES=/home/morrellp/large/soybean_toy-all-libs/All-libraries.txt
# What is the size of the genome in basepairs (for whole-genome sequencing)
# or capture region (for exome capture)?
# If unavailable, put "NA"
TARGET=NA
############################################
########## Adapter_Trimming ##########
############################################
# Which queue are we submitting the job to?
# Note: For USE_PBS, we can only specify one queue
# For USE_SLURM, we can specify multiple queues that are comma separated, no spaces (e.g., "small,ram256g,ram1t")
AT_QUEUE="small"
# USE_SLURM: What are our sbatch settings?
# Below are the recommended settings
AT_SBATCH="--nodes=1 --ntasks-per-node=4 --mem=1gb --tmp=1gb -t 10:00:00 --mail-type=ALL --mail-user=${EMAIL} -p ${AT_QUEUE}"
# USE_PBS: What are our QSub settings for Adapter_Trimming?
# Below are the recommended settings
AT_QSUB="mem=1gb,nodes=1:ppn=4,walltime=10:00:00"
# Where is the list of original FastQ files?
# Include the full file path.
RAW_SAMPLES=/home/morrellp/large/soybean_toy-all-libs/All-libraries.txt
# What shared suffix do the forward samples have?
# Example: _1_sequence.txt.gz
FORWARD_NAMING=R1_001.fastq.gz
# What shared suffix do the reverse samples have?
# Example: _2_sequence.txt.gz
REVERSE_NAMING=R2_001.fastq.gz
# If you have single-end samples, leave
# FORWARD_NAMING and REVERSE_NAMING
# filled with values that do NOT
# match your samples
# Where is the adapter file? Include the full physical file path.
ADAPTERS=/home/morrellp/large/Softwares/adapters.txt
# What is Scythe's prior contamination rate?
# Scythe's documentation suggests starting at
# 0.05 and then experimenting as needed
PRIOR=0.05
############################################
########## Quality_Trimming ##########
############################################
# Which queue are we submitting the job to?
# Note: For USE_PBS, we can only specify one queue
# For USE_SLURM, we can specify multiple queues that are comma separated, no spaces (e.g., "small,ram256g,ram1t")
QT_QUEUE="small"
# USE_SLURM: What are our sbatch settings?
# Below are the recommended settings
QT_SBATCH="--nodes=1 --ntasks-per-node=4 --mem=1gb --tmp=1gb -t 10:00:00 --mail-type=ALL --mail-user=${EMAIL} -p ${QT_QUEUE}"
# USE_PBS: What are our QSub settings for Quality_Trimming?
# Below are the recommended settings
QT_QSUB="mem=1gb,nodes=1:ppn=4,walltime=10:00:00"
# Where is our list of trimmed samples? Include the full file path.
#ADAPTED_LIST=${OUT_DIR}/Adapter_Trimming/${PROJECT}_trimmed_adapters.txt
ADAPTED_LIST=/home/morrellp/large/soybean_toy-all-libs/All-libraries-trimmed.txt
# What shared suffix do the trimmed forward samples have?
# If using the Adapter_Trimming handler,
# leave the following definition as is
FORWARD_ADAPTED=_Forward_ScytheTrimmed.fastq.gz
# What shared suffix do the trimmed reverse samples have?
# If using the Adapter_Trimming handler,
# leave the following definition as is
REVERSE_ADAPTED=_Reverse_ScytheTrimmed.fastq.gz
# What shared suffix do the trimmed single samples have?
# If using the Adapter_Trimming handler,
# leave the following definition as is
SINGLES_ADAPTED=_Single_ScytheTrimmed.fastq.gz
# If you have single-end samples, leave
# FORWARD_ADAPTED and REVERSE_ADAPTED
# filled with values that do NOT
# match your samples
# If you have paired-end samples, leave
# SINGLES_ADAPTED filled with values that
# do NOT match your samples
# What is the threshold for trimming?
# For standard trimming, use 20
QT_THRESHOLD=20
############################################
########## Read_Mapping ##########
############################################
# Which queue are we submitting the job to?
# Note: For USE_PBS, we can only specify one queue
# For USE_SLURM, we can specify multiple queues that are comma separated, no spaces (e.g., "small,ram256g,ram1t")
RM_QUEUE="small"
# USE_SLURM: What are our sbatch settings?
# Below are the recommended settings
RM_SBATCH="--nodes=1 --ntasks-per-node=16 --mem=22gb --tmp=2gb -t 24:00:00 --mail-type=ALL --mail-user=${EMAIL} -p ${RM_QUEUE}"
# USE_PBS: What are our QSub settings for Read_Mapping?
# Below are the recommended settings
# For large files, it may be necessary to use mesabi
RM_QSUB="mem=22gb,nodes=1:ppn=16,walltime=24:00:00"
# Where is our list of trimmed samples?
# Include the full file path.
TRIMMED_LIST=/home/morrellp/large/soybean_toy-all-libs/All-libraries-trimmed-2.txt
# How are trimmed forward samples named?
# If using the Adapter_Trimming handler
# FORWARD_TRIMMED=_Forward_ScytheTrimmed.fastq.gz
# If using the Quality_Trimming handler
# FORWARD_TRIMMED=_R1_trimmed.fastq.gz
FORWARD_TRIMMED=_Forward_ScytheTrimmed.fastq.gz
# How are trimmed reverse samples named?
# If using the Adapter_Trimming handler
# REVERSE_TRIMMED=_Reverse_ScytheTrimmed.fastq.gz
# If using the Quality_Trimming handler
# REVERSE_TRIMMED=_R2_trimmed.fastq.gz
REVERSE_TRIMMED=_Reverse_ScytheTrimmed.fastq.gz
# How are trimmed single-end samples named?
# If using the Adapter_Trimming handler
# SINGLES_TRIMMED=_Single_ScytheTrimmed.fastq.gz
# If using the Quality_Trimming handler
# SINGLES_TRIMMED=_single_trimmed.fastq.gz
SINGLES_TRIMMED=_Single_ScytheTrimmed.fastq.gz
# BWA mem parameters; below are the defaults for BWA mem
# Note that you may need to adjust parameters based on species
# How many threads are we using?
THREADS=16
# What is our minimum seed length?
SEED=8
# What is the band width?
WIDTH=100
# What is our off-diagonal x-dropoff (Z-dropoff)?
DROPOFF=100
# What is our re-seed value?
RE_SEED=1.0
# What is our cutoff value?
CUTOFF=10000
# What is our matching score?
MATCH=1
# What is our mismatch penalty?
MISMATCH=4
# What is our gap penalty?
GAP=8
# What is our gap extension penalty?
EXTENSION=1
# What is our clipping penalty?
CLIP=5
# What is our unpaired read penatly?
UNPAIRED=9
# Do we rescue missing hits? Note, this means that reads may not be matched. Requires paired-end mode
RESCUE=false
# Do we assume the first input query is interleaved?
INTERLEAVED=false
# What is our minimum threshold?
RM_THRESHOLD=85
# Do we output all alignments, and mark as secondary?
SECONDARY=false
# Do we append FASTA/Q comments to SAM files?
APPEND=false
# Do we use hard clipping?
HARD=false
# Do we mark split hits as secondary?
SPLIT=true
# What is the verbosity level?
# Choose from:
# 'disabled', 'errors'
# 'warnings', 'all', or 'debug'
VERBOSITY='all'
############################################
########## SAM_Processing ##########
############################################
# Which queue are we submitting the job to?
# Note: For USE_PBS, we can only specify one queue
# For USE_SLURM, we can specify multiple queues that are comma separated, no spaces (e.g., "small,ram256g,ram1t")
SP_QUEUE="small"
# USE_SLURM: What are our sbatch settings?
# Below are the recommended settings
SP_SBATCH="--nodes=1 --ntasks-per-node=16 --mem=22gb --tmp=8gb -t 24:00:00 --mail-type=ALL --mail-user=${EMAIL} -p ${SP_QUEUE}"
# USE_PBS: What are our QSub settings for SAM_Processing?
# Below are the recommended settings
SP_QSUB="mem=22gb,nodes=1:ppn=16,walltime=24:00:00"
# Specify how you would like your SAM files to be processed
# Choose from:
# 'picard' (recommended) or 'samtools'
METHOD=picard
# Number of threads used for SAM_Processing
# Using more threads requires more memory
# This is ignored when USE_PBS=true
SAM_PROCESSING_THREADS=120
# Where is the list of read-mapped samples?
# To generate this list, use sample_list_generator.sh
# located at /sequence_handling/HelperScripts/sample_list_generator.sh
MAPPED_LIST=/home/morrellp/large/soybean_toy-all-libs/All-libraries-SAM-2.txt
# The next two variables are only used if processing with Picard
# If using SAMtools, leave these variables blank
# What is the maximum number of file handles that we can use?
# For UNIX systems, the per-process maximum number of files
# that can be open may be found with 'ulimit -n'
# We recommend setting MAX_FILES to be slightly under this value
MAX_FILES=1000
# What is the maximum number (integer) of records stored in RAM before spilling to disk?
# This is used in the Picard SortSam and MarkDuplicates and is ignored if SAM_Processing with Samtools.
# You can try reducing this number if you are having core dump/out of memory errors.
# Increasing this number reduces the number of file handles needed to sort the file,
# and increases the amount of RAM needed.
# Default value: 500000
PICARD_MAX_REC_IN_RAM=500000
# What is the sorting collection size ratio we want to use?
# This is used in Picard MarkDuplicates and is ignored if SAM_Processing with Samtools.
# You can try reducing this number if you are having core dump/out of memory errors.
# Default value: 0.25
SORTING_COLL_SIZE_RATIO=0.25
############################################
########## Coverage_Mapping ##########
############################################
# Which queue are we submitting the job to?
# Note: For USE_PBS, we can only specify one queue
# For USE_SLURM, we can specify multiple queues that are comma separated, no spaces (e.g., "small,ram256g,ram1t")
CM_QUEUE="small"
# USE_SLURM: What are our sbatch settings?
# Below are the recommended settings
CM_SBATCH="--nodes=1 --ntasks-per-node=16 --mem=22gb --tmp=2gb -t 24:00:00 --mail-type=ALL --mail-user=${EMAIL} -p ${CM_QUEUE}"
# USE_PBS: What are our Qsub settings for Coverage_Mapping?
# Below are the recommended settings
CM_QSUB="mem=22gb,nodes=1:ppn=16,walltime=24:00:00"
# Where is the list of finished BAM files?
# To generate this list, use sample_list_generator.sh
# located at /sequence_handling/HelperScripts/sample_list_generator.sh
BAM_LIST=/home/morrellp/large/soybean_toy-all-libs/All-libraries-BAM.txt
# For exome capture, use the capture regions file (BED format)
# For whole-genome sequencing, leave this blank
# Coverage_Mapping should not be used for GBS data
REGIONS_FILE=
############################################
########## Haplotype_Caller ##########
############################################
# Which queue are we submitting the job to?
# Note: For USE_PBS, we can only specify one queue
# For USE_SLURM, we can specify multiple partitions that are comma separated, no spaces (e.g., "small,ram256g,ram1t")
HC_QUEUE="ram1t"
# USE_SLURM: What are our sbatch settings?
# Below are the recommended settings
HC_SBATCH="--nodes=1 --ntasks-per-node=4 --mem=250gb --tmp=22gb -t 24:00:00 --mail-type=ALL --mail-user=${EMAIL} -p ${HC_QUEUE}"
# USE_PBS: What are our Qsub settings for Haplotype_Caller?
# Below are the recommended settings
# NOTE: "mem=" is used even if qsub is not used
HC_QSUB="mem=250gb,nodes=1:ppn=24,walltime=24:00:00"
# Number of threads used for Haplotype_Caller
# Using more threads requires more memory
# This is ignored when USE_PBS=true
# Default: "NA"
HAPLOTYPE_CALLER_THREADS=120
# Where is the list of finished BAM files?
# To generate this list, use sample_list_generator.sh
# located at /sequence_handling/HelperScripts/sample_list_generator.sh
FINISHED_BAM_LIST=/home/morrellp/large/soybean_toy-all-libs/All-libraries-BAM-Chr.txt
# What is the nucleotide diversity per base pair (Watterson's theta)?
# For barley: 0.008
# For soybean: 0.001
THETA=0.001
# If true, GATK will not trim down the active region from the full region
# (active + extension) to just the active interval for genotyping
# Recommended value: false
DO_NOT_TRIM_ACTIVE_REGIONS=false
# If true, all bases will be considered active regions
# Recommended value: false
FORCE_ACTIVE=false
# Optional: Provide a list of custom intervals if you are interested in specific regions AND/OR want to parallelize processing across regions/chromosomes.
# Default: false. Otherwise, fill in full path to intervals file.
# IMPORTANT: GATK accepts the following file extensions:
# .intervals, .list, .interval_list, .bed, and .vcf
# sequence_handling Haplotype_Caller currently handles: .intervals, .list
# For .intervals/.list files, format one per line in one of the two formats: 1) chr1 or 2) chr1:100-200
# For example files, see: https://github.com/MorrellLAB/sequence_handling/tree/master/Example_Files
# For additional info, please see Wiki page.
HC_CUSTOM_INTERVALS=false
# Optional: If your genome has scaffolds or parts of the reference not covered by
# the chromosomes above, include the full filepath to a SORTED list of
# names. One scaffold name per line.
# IMPORTANT: If you provide a scaffolds list, you must provide a custom intervals list too.
# Default is set to false. Otherwise, fill in full path to intervals file.
HC_SCAFFOLDS=false
# Do we want to parallelize across regions?
# Set to true if yes.
# Default is set to false.
# Parallelizing across regions for Haplotype_Caller means each
# region/chromosome provided in HC_CUSTOM_INTERVALS list gets run
# in as it's own job for every sample.
HC_PARALLELIZE=false
################################################
########## Genomics_DB_Import ##########
################################################
# USAGE NOTE: Genomics DB Import is used in GATK4.
# Which queue are we submitting the job to?
# Note: For USE_PBS, we can only specify one queue
# For USE_SLURM, we can specify multiple queues that are comma separated, no spaces (e.g., "small,ram256g,ram1t")
GDBI_QUEUE="amdsmall,amd512,amd2tb"
# USE_SLURM: What are our sbatch settings?
# Below are the recommended settings
GDBI_SBATCH="--nodes=1 --ntasks-per-node=4 --mem=248gb --tmp=100gb -t 96:00:00 --mail-type=ALL --mail-user=${EMAIL} -p ${GDBI_QUEUE}"
# USE_PBS: What are our Qsub settings for Genomics_DB_Import?
# Below are the default settings but this depends on the number of samples
# NOTE: mem has to be in the unit of gb, and it has to be greater than 4gb
# NOTE2: "mem=" is used even if qsub is not used
GDBI_QSUB="mem=400gb,nodes=1:ppn=4,walltime=96:00:00"
# Number of threads used for Genomics_DB_IMPORT
# Using more threads requires more memory
# This is ignored when USE_PBS=true
GDBI_THREADS=120
# REQUIRED: Please fill out variables under "Genotype_GVCFs" listed below.
# Genomics_DB_Import shares variables with Genotype_GVCFs.
############################################
########## Genotype_GVCFs ##########
############################################
# USAGE NOTE: For accurate results, run Genotype_GVCFs on all of your samples at once
# DO NOT break this job into batches of samples
# Which queue are we submitting the job to?
# Note: For USE_PBS, we can only specify one queue
# For USE_SLURM, we can specify multiple queues that are comma separated, no spaces (e.g., "small,ram256g,ram1t")
GG_QUEUE="amdsmall,amd512,amd2tb"
# USE_SLURM: What are our sbatch settings?
# Below are the recommended settings
GG_SBATCH="--nodes=1 --ntasks-per-node=4 --mem=248gb --tmp=100gb -t 96:00:00 --mail-type=ALL --mail-user=${EMAIL} -p ${GG_QUEUE}"
# USE_PBS: What are our Qsub settings for Genotype_GVCFs?
# Below are the recommended settings
# NOTE: "mem=" is used even if qsub is not used
GG_QSUB="mem=248gb,nodes=1:ppn=4,walltime=96:00:00"
# Number of threads used for Genotype_GVCFs
# Using more threads requires more memory
# This is ignored when USE_PBS=true
GG_THREADS=120
# Where is the list of GVCF files?
# To generate this list, use sample_list_generator.sh
# located at /sequence_handling/HelperScripts/sample_list_generator.sh
GVCF_LIST=/home/morrellp/large/soybean_toy-all-libs/All-libraries-VCF.txt
# What is the nucleotide diversity per base pair (Watterson's theta)?
# Genotype_GVCFs uses the THETA under Haplotype_Caller for this
# Where is the reference dictionary file? (.dict)
# Include the full file path.
REF_DICT=/home/morrellp/large/soybean_toy/genome/Gmax_275_v2.0.dict
# How many chromosomes or chromosome parts does the reference have, excluding scaffolds? (integer value)
# For barley: 15 (7*2 chromosome parts + chrUn)
# For soybean: 20
NUM_CHR=20
# Optional: Provide a list of custom intervals if you are interested in specific regions AND/OR want to parallelize processing across regions/chromosomes.
# IMPORTANT: GATK accepts the following file extensions:
# .intervals, .list, .interval_list, .bed, and .vcf
# sequence_handling currently handles: .intervals, .list, and .bed
# For .intervals/.list files, format one per line as follows: chr1:100-200
# For example files, see: https://github.com/MorrellLAB/sequence_handling/tree/master/Example_Files
# For additional info, please see Wiki page.
# Default is set to false. Otherwise, fill in full path to intervals file.
# NOTE: If HC_PARALLELIZE=true and HC_CUSTOM_INTERVALS is set, the value below is ignored (uses HC_CUSTOM_INTERVALS).
CUSTOM_INTERVALS=/home/morrellp/large/soybean_toy/genome/Gmax_275_v2.0-Chr.list
# Optional: If your genome has scaffolds or parts of the reference not covered by
# the chromosomes above, include the full filepath to a SORTED list of
# names. One scaffold name per line.
# Default is set to false. Otherwise, fill in full path to intervals file.
# NOTE: If HC_PARALLELIZE=true and HC_CUSTOM_INTERVALS is set, the value below is ignored (uses HC_SCAFFOLDS).
SCAFFOLDS=/home/morrellp/large/soybean_toy/genome/scaffolds.intervals
#SCAFFOLDS=
# Do we want to parallelize across regions or chromosomes?
# Set to true if yes.
# Default is set to false.
# Parallelizing across regions means each region/chromosome provided in
# CUSTOM_INTERVALS list gets run as its own job.
# In the case that we are parallelizing across regions AND we have scaffolds,
# regions in CUSTOM_INTERVALS will be parallelized but the scaffolds will not be parallelized.
# Scaffolds will instead be imported into its own gendb workspace.
# NOTE: If HC_PARALLELIZE=true and HC_CUSTOM_INTERVALS is set, the value below is ignored.
PARALLELIZE=true
# When parallelized, separate vcf output files are created for separate intervals in
# ${OUT_DIR}/Genotype_GVCFs/vcf_split_regions
# If you set the following option to true, these separate files get combined into a single vcf:
# ${OUT_DIR}/Genotype_GVCFs/raw_variants.vcf
# This option is ignored if PARALLELIZE=false
# NOTE: this could require lots of memory, so if it fails, you need to increase
# mem= option in GG_QSUB (this option is used even if you are not using qsub)
# IMPORTANT: Currently, this option only works for non-PBS. For PBS users, split VCF files will
# get combined and sorted in the Create_HC_Subset handler. If you don't plan to use the
# Create_HC_Subset handler, the HelperScripts/combine_and_sort_split_vcf.sh
# script will combine and sort the split VCF files for you.
# Default is set to true.
GG_COMBINED_VCF=true
# What is the sample ploidy? (integer value)
# For highly inbred samples (most barleys): 1
PLOIDY=2
############################################
########## Create_HC_Subset ##########
############################################
# Which queue are we submitting the job to?
# Note: For USE_PBS, we can only specify one queue
# For USE_SLURM, we can specify multiple queues that are comma separated, no spaces (e.g., "small,ram256g,ram1t")
CHS_QUEUE="amdsmall,amd512,amd2tb"
# USE_SLURM: What are our sbatch settings?
# Below are the recommended settings
CHS_SBATCH="--nodes=1 --ntasks-per-node=16 --mem=22gb --tmp=8gb -t 24:00:00 --mail-type=ALL --mail-user=${EMAIL} -p ${CHS_QUEUE}"
# USE_PBS: What are our Qsub settings for Create_HC_Subset?
# Below are the recommended settings
CHS_QSUB="mem=22gb,nodes=1:ppn=16,walltime=24:00:00"
# Where is our raw variants VCF file?
# If we parallelized across regions in Genotype_GVCFs and don't have a
# concatenated VCF file (GG_COMBINED_VCF was set to false), fill out the next variable
# called "CHS_VCF_LIST=" with the full filepath to a list of split vcf files.
# IMPORTANT: DO NOT LEAVE BLANK. If you don't have a single raw variants VCF file and are going to concatenate split VCF files
# put "NA" here. Odd errors may appear if this is not filled out.
CHS_RAW_VCF="NA"
# Where is the list of chromosome part VCF files?
# If this is filled out, this handler will concatenate VCF files split into parts.
# NOTE: This concatenation and sorting feature requires lots of memory, so keep adding more memory if you get out of memory errors.
# To generate this list, use sample_list_generator.sh
# located at /sequence_handling/HelperScripts/sample_list_generator.sh
# Default is "NA", replace with full filepath to list of split VCF files if you do not have a single raw variants VCF
CHS_VCF_LIST=/home/morrellp/large/soybean_toy-all-libs/All-libraries-VCF-genotyped.txt
# If exome capture, include the full file path to the capture regions file (BED format)
# This should be the same file as the REGIONS_FILE in Coverage_Mapping. In this case, put "${REGIONS_FILE}" below.
# If not exome capture, put "NA"
CAPTURE_REGIONS="NA"
# Subset unfiltered vcf to one chromosome for percentile tables used to get a sense of the distirbution of per sample DP and GQ?
# Recommended for very large genomes where too much memory is required and job fails due to running out of memory
# Default is "NA", replace with exact matching chromosome name. For more than one chromosome (or chromosome part), use comma WITHOUT spaces to delimit. Example: "chr1H_part1,chr1H_part2"
SUBSET_CHR="NA"
# File prefix we should use for subsetted output file
# Default is "NA". Example: "chr1H"
SUBSET_CHR_OUT_PREFIX="NA"
# What is the depth per sample (DP) cutoff? (integer)
# If a sample's DP is below this threshold, it will count as a "bad" sample for that site.
# Recommended value: 5
CHS_DP_PER_SAMPLE_CUTOFF=5
# What is the genotyping quality (GQ) cutoff? (integer)
# If a sample's GQ is below this threshold, it will count as a "bad" sample for that site.
# Recommended value: 10th percentile of the raw GQ percentile table
CHS_GQ_CUTOFF=10
# What is the maximum proportion of "lowGQ" (below the GQ cutoff) samples allowed per site?
# Sites with more "lowGQ" samples than this threshold will be filtered out.
# Recommended value: "0.2"
CHS_MAX_LOWGQ=0.2
# What is the maximum proportion of heterozygous genotypes per site? (proportion)
# Sites with a proportion above this threshold will be filtered out
# Recommended value: "0.1" based on highly inbred species like barley
CHS_MAX_HET=0.1
# What is the maximum proportion of missing calls per site? (proportion)
# Sites with a proportion above this threshold will be filtered out
# Recommended value: "0.2"
CHS_MAX_MISS=0.2
# What is the QUAL score cutoff? (integer)
# Sites with a QUAL below this cutoff will be excluded.
# Recommended value: 40
CHS_QUAL_CUTOFF=40
# How much of the variants do you want to sample randomly for QC graphs (as genome intervals)?
# The number of genomic regions to sample:
CHS_SUBSET_N=1000000
# The length (in base pairs) of genomic regions to sample:
CHS_SUBSET_LEN=100
############################################
########## Variant_Recalibrator ##########
############################################
# DO NOT RUN VARIANT_RECALIBRATOR IF YOU HAVE LESS THAN 30 EXOME SAMPLES
# GATK has empirically found in human data that you need at least 1 whole genome or 30 exome samples to have enough variant sites for decent modeling.
# What matters is having a large number of variant sites, so this minimum number of samples is only an estimate.
# See documentation here for details: https://gatk.broadinstitute.org/hc/en-us/articles/360035531612-Variant-Quality-Score-Recalibration-VQSR-
# Which queue are we submitting the job to?
# Note: For USE_PBS, we can only specify one queue
# For USE_SLURM, we can specify multiple queues that are comma separated, no spaces (e.g., "small,ram256g,ram1t")
VR_QUEUE="amdlarge,amdsmall,amd512,amd2tb"
# USE_SLURM: What are our sbatch settings?
# Below are the recommended settings
VR_SBATCH="--nodes=1 --ntasks-per-node=4 --mem=240gb --tmp=22gb -t 24:00:00 --mail-type=ALL --mail-user=${EMAIL} -p ${VR_QUEUE}"
# USE_PBS: What are our Qsub settings for Variant_Recalibrator?
# Below are the recommended settings
VR_QSUB="mem=240gb,nodes=1:ppn=4,walltime=24:00:00"
# What reference genome are we using?
# Include the full file path.
# If barley AND running GATK3, make sure to use the pseudomolecular reference
# NOT the parts reference used above
# If running GATK4, make sure this is the same reference used during Haplotype_Caller and Genotype_GVCFs
VR_REF=/home/morrellp/large/soybean_toy/genome/Gmax_275_v2.0.fa
# Do we have a single concatenated raw VCF file output from Create_HC_Subset?
# If not, put "NA" below
# GATK4 only accepts a single input variant file unlike earlier version of GATK, which accepted multiple input variant files.
# To generate this list, use sample_list_generator.sh
# located at /sequence_handling/HelperScripts/sample_list_generator.sh
VR_RAW_VCF="NA"
# Where is the list of chromosome part VCF files from Genotype_GVCFs?
# Leave as the default "NA" if we already have a concatenated raw VCF file. Change otherwise.
# GATK3 accepts multiple input variant files but GATK4 does not. If we are running GATK4, the handler will
# take the list of split VCF files and concatenate them for you.
# To generate this list, use sample_list_generator.sh
# located at /sequence_handling/HelperScripts/sample_list_generator.sh
VR_VCF_LIST=/home/morrellp/large/soybean_toy-all-libs/All-libraries-VCF-genotyped.txt
# Do we want to recalibrate indels, SNPs, or both?
# Default is "BOTH"
# Specifically, indels are recalibrated first then SNPs as recommended by GATK best practices
# Valid values (case sensitive) include: "BOTH", "INDELS_ONLY", "SNPS_ONLY"
# "BOTH" means indels are recalibrated first then SNPs as recommended by GATK best practices
# "INDELS_ONLY" pulls out indels and only recalibrates indels
# "SNPS_ONLY" pulls out SNPs and only recalibrates SNPs
RECAL_MODE="BOTH"
# Which annotations do we want to include?
# INDEL and SNPs are recalibrated separately, for flexibility you can specify different sets of annotations to use for each below.
# Specifically, read up on GATK Variant Recalibrator's caveats for specific types of data (example: DP annotation should not be used for exome datasets)
# See doc here for what the annotations mean: https://gatk.broadinstitute.org/hc/en-us/articles/360035890471
# NOTE: MQ is intentionally left out when recalibrating indels, see this tutorial for more info: https://gatk.broadinstitute.org/hc/en-us/articles/360035531112--How-to-Filter-variants-either-with-VQSR-or-by-hard-filtering
# IMPORTANT: DP should NOT be used for exome datasets (see here https://gatk.broadinstitute.org/hc/en-us/articles/360035531612-Variant-Quality-Score-Recalibration-VQSR-)
# DEFAULT annotations are included below, please add/exclude annotations as you see fit.
# Please following this example format to add/exlude annotations:
# VR_ANN_INDEL="-an FS -an ReadPosRankSum -an MQRankSum -an QD -an SOR -an DP"
VR_ANN_INDEL="-an QD -an FS -an ReadPosRankSum -an MQRankSum -an SOR -an DP"
VR_ANN_SNP="-an QD -an FS -an ReadPosRankSum -an MQ -an MQRankSum -an SOR -an DP"
# What truth sensitivity tranche settings do we want to use?
# These are the levels of truth sensitivity at which to slice the data (in percent)
# and will be used with the VariantRecalibrator function call.
# If modifying, please follow the format: "-tranche 100.0 -tranche 99.0"
TRANCHE_INDEL="-tranche 100.0 -tranche 99.9 -tranche 99.0 -tranche 97.0 -tranche 96.0 -tranche 95.0 -tranche 94.0 -tranche 93.0 -tranche 92.0 -tranche 91.0 -tranche 90.0"
TRANCHE_SNP="-tranche 100.0 -tranche 99.95 -tranche 99.9 -tranche 99.8 -tranche 99.6 -tranche 99.5 -tranche 99.4 -tranche 99.3 -tranche 99.0 -tranche 98.0 -tranche 97.0 -tranche 90.0"
# EXTRA FLAGS. Are there additional flags (options) you would like to include?
# Default is "NA", modify otherwise. DO NOT leave blank!
# Use the same options format as listed in the gatk docs except separated by space, all on one line, and surrounded by double quotes.
# Example format: RECAL_EXTRA_OPTIONS_INDEL="-L genomic_intervals -titv 2.15"
# Recalibration step:
# INDEL mode: Currently, indel recalibration uses the following flags:
# -R, -V, -an (see VR_ANN_INDEL above), -tranche (see TRANCHE_INDEL above), -mode INDEL, -O, --resource, --tranches-file, --rscript-file
RECAL_EXTRA_OPTIONS_INDEL="NA"
# SNP mode: Currently, SNP recalibration uses the following flags:
# -R, -V, -an (see VR_ANN_SNP above), -tranche (see TRANCHE_SNP above), -mode SNP, -O, --resource, --tranches-file, --rscript-file
RECAL_EXTRA_OPTIONS_SNP="NA"
# ApplyVQSR step (filtering step):
# INDEL mode: Currently, indel ApplyVQSR uses the following options:
# -R, -V, -mode INDEL, --truth-sensitivity-filter-level, --recal-file, --tranches-file, --create-output-variant-index, -O
FILTER_EXTRA_OPTIONS_INDEL="NA"
# SNP mode: Currently, snp ApplyVQSR uses the following options:
# -R, -V, -mode SNP, --truth-sensitivity-filter-level, --recal-file, --tranches-file, --create-output-variant-index, -O
FILTER_EXTRA_OPTIONS_SNP="NA"
# Where are resource VCF files and their priors (integers) to be used for training our model?
# At least one resource (high confidence subset counts as one) and prior pair is required.
# MPORTANT: GATK 4 requires both training AND truth sets to run, otherwise it will return an error.
# These can be supplied in a single resource or as separate resources (fill out the next section below).
# Where is the high confidence subset of variants?
# The high confidence set recommended settings: known=false,training=true,truth=false
# If you used Create_HC_Subset, leave as the default
HC_SUBSET=${OUT_DIR}/Create_HC_Subset/${PROJECT}_high_confidence_subset.vcf
HC_PRIOR=5.0
HC_KNOWN=false
HC_TRAIN=true
HC_TRUTH=false
# Do we have additional resource VCF files we can use?
# "NA" is the default for missing resource files and priors, modify otherwise
# Please specify "true" or "false" for KNOWN, TRAINING, and TRUTH for each resource below (this is currently set to defaults)
# See GATK's documentation for definitions of known, training, and truth sets:
# https://gatk.broadinstitute.org/hc/en-us/articles/360035890831-Known-variants-Training-resources-Truth-sets
RESOURCE_1=/home/morrellp/large/soybean_toy/genome/training_VCF_files/M92-220_final_filtered-edited.vcf
PRIOR_1=5.0
KNOWN_1=false
TRAINING_1=false
TRUTH_1=true
RESOURCE_2=/home/morrellp/large/soybean_toy/genome/training_VCF_files/BL107_final_filtered.vcf
PRIOR_2=5.0
KNOWN_2=false
TRAINING_2=false
TRUTH_2=true
RESOURCE_3=/home/morrellp/large/soybean_toy/genome/training_VCF_files/50k-SNPs-sorted-SNP_utils.vcf
PRIOR_3=5.0
KNOWN_3=true
TRAINING_3=true
TRUTH_3=false
RESOURCE_4="NA"
PRIOR_4="NA"
KNOWN_4=true
TRAINING_4=true
TRUTH_4=false
# What is our truth sensitivity filter level?
# Default is set to 99.9
TS_FILTER_LEVEL="99.9"
################################################
########## Pre_Variant_Filtering ##########
################################################
# Which queue are we submitting the job to?
# Note: For USE_PBS, we can only specify one queue
# For USE_SLURM, we can specify multiple queues that are comma separated, no spaces (e.g., "small,ram256g,ram1t")
PVF_QUEUE="amdsmall,amd512,amd2tb"
# USE_SLURM: What are our sbatch settings?
# Below are the recommended settings
PVF_SBATCH="--nodes=1 --ntasks-per-node=16 --mem=110gb --tmp=22gb -t 72:00:00 --mail-type=ALL --mail-user=${EMAIL} -p ${PVF_QUEUE}"
# USE_PBS: What are our Qsub settings?
# Below are the recommended settings
PVF_QSUB="mem=110gb,nodes=1:ppn=16,walltime=72:00:00"
# Where is the VCF file we can use to graph annotations and generate percentile tables?
# If you ran Variant_Recalibrator, this will be the .recalibrated.vcf.gz file
PVF_VCF=/home/morrellp/large/soybean_toy-all-libs/Variant_Recalibrator/soybean_toy_dataset_all_libs_indels_and_snps.recalibrated.vcf.gz
# What is the number of genomic regions to subset for graphing annotation purposes?
# Default: 1000000
PVF_GEN_NUM="1000000"
# What is the length of genomic regions to subset for graphing annotation purposes?
# Default: 100
PVF_GEN_LEN="100"
# NOTE:
# This handler works with the GATK VariantsToTable function output.
# See more details about what each annotation means here:
# https://gatk.broadinstitute.org/hc/en-us/articles/360036732151-VariantsToTable
# https://gatk.broadinstitute.org/hc/en-us/articles/360037261311-ExcessHet
# https://gatk.broadinstitute.org/hc/en-us/articles/360036366852-InbreedingCoeff
############################################
########## Variant_Filtering ##########
############################################
# Which queue are we submitting the job to?
# Note: For USE_PBS, we can only specify one queue
# For USE_SLURM, we can specify multiple queues that are comma separated, no spaces (e.g., "small,ram256g,ram1t")
VF_QUEUE="amdsmall,amd512,amd2tb"
# USE_SLURM: What are our sbatch settings?
# Below are the recommended settings
VF_SBATCH="--nodes=1 --ntasks-per-node=16 --mem=110gb --tmp=22gb -t 72:00:00 --mail-type=ALL --mail-user=${EMAIL} -p ${VF_QUEUE}"
# USE_PBS: What are our Qsub settings?
# Below are the recommended settings
VF_QSUB="mem=110gb,nodes=1:ppn=16,walltime=72:00:00"
# Where is the VCF file to be filtered?
# If you ran Variant_Recalibrator and Pre_Variant_Filtering, this will be
# the .recalibrated.pass_sites.vcf.gz file
VF_VCF=/home/morrellp/large/soybean_toy-all-libs/Pre_Variant_Filtering/soybean_toy_dataset_all_libs_indels_and_snps.recalibrated.pass_sites.vcf.gz
# What is the QUAL score cutoff? (integer)
# Sites with a QUAL below this cutoff will be excluded.
# Recommended value: 40
VF_QUAL_CUTOFF=40
# What is the minimum number of reads needed to support a genotype? (integer)
# Recommended value: 5
VF_MIN_DP="5"
# What is the maximum number of reads allowed to support a genotype? (too many = gene duplication problems) (integer)
# You can use the Pre_Variant_Filtering annotation plots and DP per sample percentile table as a guide, but please do
# to decide on a final cutoff.
# Default value: 100 (but please adjust accordingly for your study!)
VF_MAX_DP="100"
# What is the genotyping quality (GQ) cutoff? (integer)
# If a sample's GQ is below this threshold, it will get filtered out
# Recommended value: 10th percentile of the recal_pass_sites GQ percentile table
# Example format: 3
VF_GQ_CUTOFF="3"
# What is the QD cutoff?
# Sites below this cutoff will be filtered out.
# See documentation for details: https://gatk.broadinstitute.org/hc/en-us/articles/360036732151-VariantsToTable
VF_QD_CUTOFF="2.0"
# What is the SOR cutoff?
# Sites above this cutoff will get filtered out.
# See documentation for details: https://gatk.broadinstitute.org/hc/en-us/articles/360036732151-VariantsToTable
VF_SOR_CUTOFF="3.0"
# What is the FS cutoff?
# Sites above this cutoff will get filtered out.
# See documentation for details: https://gatk.broadinstitute.org/hc/en-us/articles/360036732151-VariantsToTable
VF_FS_CUTOFF="60.0"
# What is the MQ cutoff?
# Sites below this cutoff will get filtered out.
# See documentation for details: https://gatk.broadinstitute.org/hc/en-us/articles/360036732151-VariantsToTable
VF_MQ_CUTOFF="40.0"
# What is the proportion of missingness we are willing to tolerate? (proportion)
# Sites with a misssingness greater than this threshold will be filtered out
# Default value: "0.25"
VF_MISS="0.25"
# What is the maximum proportion of heterozygous genotypes allowed per site? (proportion)
# Sites with a proportion above this threshold will be filtered out
# Recommended value: "0.1" based on highly inbred species like barley
VF_MAX_HET="0.1"
# What is the number of genomic regions to subset for graphing annotation purposes?
# Default: 1000000
VF_GEN_NUM="1000000"
# What is the length of genomic regions to subset for graphing annotation purposes?
# Default: 100
VF_GEN_LEN="100"
#----------------
# The following filters have caveats and will only be used when filtering if the conditions are met
# However, they must be filled out!
# Are the samples diploid?
# Valid options are: 1) "true" or 2) "false"
# Default: false
VF_DIPLOID="false"
# If we have diploid samples, what excess heterozygosity cutoff should we use? (integer)
# See documentation for details: https://gatk.broadinstitute.org/hc/en-us/articles/360037261311-ExcessHet
# Refer to the Pre_Variant_Filtering plots to determine an appropriate cutoff.
# Caveat: Can only be calculated for diploid samples.
# Default: "NA"
VF_EXCESS_HET="NA"
#-----------------
### GATK3 specific variables for Variant_Filtering handler
# If GATK4 was used, delete the following GATK3 specific variables in the Variant_Filtering handler
# What is the maximum percent deviation from 50/50 reference/alternative reads allowed in heterozygotes? (decimal)
# For example, MAX_DEV=0.1 allows 60/40 ref/alt and also 40/60 ref/alt but not anything more skewed
# Recommended value: 0.1
VF_MAX_DEV=0.1
# What is the maximum number of "bad" (low GQ, low DP, or missing genotype data) samples allowed at a site? (integer)
# Sites with more "bad" samples than this threshold will be filtered out.
# Recommended value: "0.2"
VF_MAX_BAD=0.2
# If exome capture, include the full file path to the capture regions file (BED format)
# For barley use the pseudomolecular BED file, not the parts file
# If not exome capture, put "NA"
# Note: This is specific to GATK 3. If you are running GATK 4, you can leave this blank.
VF_CAPTURE_REGIONS="NA"
#-----------------
############################################
########## Variant_Analysis ##########
############################################
# Which queue are we submitting the job to?
# Note: For USE_PBS, we can only specify one queue
# For USE_SLURM, we can specify multiple queues that are comma separated, no spaces (e.g., "small,ram256g,ram1t")
VA_QUEUE="amdsmall,amd512,amd2tb"
# USE_SLURM: What are our sbatch settings?
# Below are the recommended settings
VA_SBATCH="--nodes=1 --ntasks-per-node=16 --mem=22gb --tmp=4gb -t 24:00:00 --mail-type=ALL --mail-user=${EMAIL} -p ${VA_QUEUE}"
# What are our Qsub settings for Variant_Analysis?
# Below are the recommended settings
VA_QSUB="mem=22gb,nodes=1:ppn=16,walltime=24:00:00"
# Where is the VCF file? Include the full file path.
VA_VCF=/home/morrellp/large/soybean_toy-all-libs/Variant_Filtering/soybean_toy_dataset_all_libs_final.vcf.gz
# Do we have a Sanger dataset we can make comparisons to?
# If so, fill out the full filepath to the Sanger regions in BED file format.
# IMPORTANT: This variable must be filled out, if you are not using these variables put "NA"
# Default: "NA"
SANGER_BED="NA"
############################################
########## Dependencies ##########
############################################