fq2bam performs the following steps. User can decide to turn-off marking of duplicates. BQSR step is only performed if
--knownSites input and
--out-recal-file options are provided.
$ pbrun fq2bam --ref Ref/Homo_sapiens_assembly38.fasta \--in-fq S1_1.fastq.gz S1_2.fastq.gz \--knownSites Ref/Homo_sapiens_assembly38.known_indels.vcf.gz \--out-bam mark_dups_gpu.bam \--out-recal-file recal_gpu.txt \--tmp-dir /raid/myrun
The command below is the bwa-0.7.15 and GATK4 counterpart of the Parabricks command above. The output from these commands will generate the exact same results as the output from the above command. Please look at Output Comparison page on how you can compare the results.
# Run bwa-mem and pipe output to create sorted bam$ bwa mem -t 32 -K 10000000 -R '@RG\tID:sample_rg1\tLB:lib1\tPL:bar\tSM:sample\tPU:sample_rg1' \Ref/Homo_sapiens_assembly38.fasta S1_1.fastq.gz S1_2.fastq.gz | gatk \SortSam --java-options -Xmx30g --MAX_RECORDS_IN_RAM=5000000 -I=/dev/stdin \-O=cpu.bam --SORT_ORDER=coordinate --TMP_DIR=/raid/myrun# Mark Duplicates$ gatk MarkDuplicates --java-options -Xmx30g -I=cpu.bam -O=mark_dups_cpu.bam \-M=metrics.txt --TMP_DIR=/raid/myrun# Generate BQSR Report$ gatk BaseRecalibrator --java-options -Xmx30g --input mark_dups_cpu.bam --output \recal_cpu.txt --known-sites Ref/Homo_sapiens_assembly38.known_indels.vcf.gz \--reference Ref/Homo_sapiens_assembly38.fasta
The reference genome in fasta format. We assume that the indexing required to run bwa has been completed by the user.
Pair ended fastq files. These can be in
This option can be repeated multiple times as well. Example 1:
Either all sets of inputs have read group or none should have it and will be automatically added by the pipeline. For same sample, Read Groups should have same sample name (SM) and different ID and PU.
Path to the file that will contain BAM output.
Path of report file (
Path of duplicate metrics file after Marking Duplicates.
Run MergeBAMAlignment after alignment.
Path to a file containing options for MergeBAMAlignment. Currently supported options are
Known indel files in
Do not mark duplicates, generate bam after co-ordinate sorting.
Pass supported bwa mem options as one string. Current
original bwa mem supported options:
Assume the reads are sorted by queryname for Marking Duplicates. This will mark secondary, supplementary and unmapped reads as duplicates as well. This flag will not impact variant calling while increasing processing times.
The maximum offset between two duplicate clusters in order to consider them optical duplicates. This option is only used if you provide an
SM tag for read groups in this run.
LB tag for read groups in this run.
PL tag for read groups in this run.
Prefix for ID and PU tag for read groups in this run. This prefix will be used for all pair of fastq files in this run. The ID and PU tag will consist of this prefix and an identifier which will be unique for a pair of fastq files.
Full path to the directory where temporary files will be stored.
Defaults to number of GPUs in the system.
The number of GPUs to be used for this analysis task.