This workflow supports the analysis of canine sequencing samples against the CanFam3.1 reference genome
using ensembl version 98 gene models. The workflow is designed to support project level analysis
that can include one or multiple types of data. Though not required the expectation is a project contains
data from a single individual thus centralizing all data types in a standardized output structure. The
workflow template supports a diverse array of analysis methods required to analyze DNA, RNA.
Based on standardized variables it also supports integrated analysis between data types. For some
processes multiple options are provided that can be individually enabled or disabled by configuration
parameters. Like all JetStream (https://github.com/tgen/jetstream) workflows developed at TGen this workflow is designed to facilitate our dynamic and
time sensitive analysis needs while ensuring compute and storage resources are used efficiently. The primary
input is a JSON record from the TGen LIMS but hand created inputs in the form of a JSON or EXCEL worksheet can
also be provided when run manually or by submission to the related JetStream Centro (https://github.com/tgen/jetstream_centro)
web portal. All required files defined the the pipeline.yaml can be created using code provided in the JetStream Resources
repository (https://github.com/tgen/jetstream_resources).
Click to show details
DNA Alignment
The DNA Alignment module supports the generation of processed BAM or CRAM files from input fastqs files.
The configuration files define one of three different alignment style variables (dnaAlignmentStyle) that
all use BWA as the aligner but use different tools for alignment processing. Additionally, baserecalibration
and/or conversion to CRAM for archiving can be enabled for any project.
- TGen [
tgen] (default)- Fastq files are chunked to 40M reads
- Chunks are aligned with BWA, processed with samtools fixmate and samtools sort
- Individual chunks are merged with samtools merge
- PCR duplicates and platform (optical) duplicates are marked with samtools markdup
- GATK Best Paractices [
broad]("Like" - does not support uBAM inputs) NOT TESTED FOR COYOTE- Fastq files are chunked to 40M reads
- Chunks are aligned with BWA, and converted to BAM files with samtools view
- In one step individual chunks are merged and PCR or platform (optical) duplicates are marked with GATK/Picard markduplicates
- Alignment records are sorted using GATK/Picard SortSAM
- Clinical [
ashion] NOT TESTED FOR COYOTE- Fastq files are concatenated
- In one step reads are aligned with BWA, duplicates are marked with samblaster, converted to BAM and sorted using sambamba
- Base Recalibration (optional, off by default)
- chunked baserecalibration using GATK
- BAM to CRAM (enabled by default)
- samtools view
Constitutional Analysis
These modules support the discovery of small variants (SNV/INDELs), structural abnormalities (deletions, duplications, inversions, translocations), and copy number abnormalities on single constitutional samples
- Variant Discovery (SNV/INDEL)
- Deepvariant (Only calls on primary contigs 1-38, X)
- requires a GPU compute resource
- GATK HaplotypeCaller (gVCF mode)
- gVCF generation
- single sample calling using genotypeGVCFs and CNNscroreVariant
- Freebayes
- Strelka2 (Germline mode)
- Octopus (Individual) (Only calls on primary contigs 1-38, X)
- Deepvariant (Only calls on primary contigs 1-38, X)
- Structural Variant Detection
- Manta
- Copy Number Analysis
- iCHOR (operates on genomes only)
Somatic Analysis
This module will run several somatic variant callers on tumor/normal data pairs:
- Variant Discovery (SNV/INDEL)
- Mutect2
- Lancet
- Currently NOT enabled for genomes by default
- VarDictJava
- Octopus (Only calls on primary contigs 1-38, X)
- Structural Variant Detection
- Manta
- Delly
- Copy Number Analysis
- GATK CNV
- iCHOR (operates on genomes only)
Tumor Only Analysis
This module will run several variant callers using a tumor sample and user defined normal data pairs:
- Variant Discovery (SNV/INDEL)
- Strelka2 (Somatic mode)
- Mutect2
- Lancet
- Currently NOT enabled for genomes by default
- VarDictJava
- Octopus (Only calls on primary contigs 1-22, X, Y)
- Structural Variant Detection
- Manta
- Delly
- Copy Number Analysis
- GATK CNV
- iCHOR (operates on genomes only)
RNA Analysis
- RNA Alignment
- STAR
- 2-pass mode
- outputs unsorted BAM by default
- option to output TranscriptomeSAM
- unsorted BAM is processed with samtools fixmate and samtools sort to output a sorted BAM
- PCR duplicates and platform duplicates (optical) are optionally marked as a QC step
- samtools markdup
- GATK/Picard MarkDuplicates (default recommendation)
- STAR
- Gene and Transcript Expression Estimates
- Salmon
- Star-count - Recommended count method
- HtSeq
- FeatureCounts
- Fusion Transcript Detection
- Star-Fusion (Currently non-functional)
Single Cell RNA Sequencing
NOT TESTED FOR COYOTE
- 10x Genomics Cell Ranger
- Supports 3' and 5' assays
- Supports VDJ analysis
- starSOLO
- Supports 3' and 5' assays
All final output files are placed in a standardized folder structure that generally reflects the relationship of files or the processing order.
Project
|--GeneralLibaryType
| |--AnalysisType
| | |--Tool
| | | |--SampleName
| | | |--ResultFiles
| | |--Tool
| |--AnalysisType
|--GeneralLibaryType
Project Folder Example
# Only Directories are Shown
MMRF_1499
├── exome
│  ├── alignment
│  │  └── bwa
│  │  ├── MMRF_1499_1_BM_CD138pos_T2_KAS5U
│  │  │  └── stats
│  │  └── MMRF_1499_2_PB_Whole_C7_KHS5U
│  │  └── stats
│  ├── constitutional_structural_calls
│  │  └── manta
│  │  └── MMRF_1499_2_PB_Whole_C7_KHS5U
│  ├── constitutional_variant_calls
│  │  ├── deepvariant
│  │  │  └── MMRF_1499_2_PB_Whole_C7_KHS5U
│  │  └── haplotypecaller
│  │  └── MMRF_1499_2_PB_Whole_C7_KHS5U
│  ├── somatic_copy_number
│  │  └── gatk
│  │  └── MMRF_1499_2_PB_Whole_C7_KHS5U-MMRF_1499_1_BM_CD138pos_T2_KAS5U
│  ├── somatic_structural_calls
│  │  ├── manta
│  │  │  └── MMRF_1499_2_PB_Whole_C7_KHS5U-MMRF_1499_1_BM_CD138pos_T2_KAS5U
│  │  └── pairoscope
│  │  └── MMRF_1499_1_BM_CD138pos_T2_KAS5U
│  └── somatic_variant_calls
│  ├── lancet
│  │  └── MMRF_1499_2_PB_Whole_C7_KHS5U-MMRF_1499_1_BM_CD138pos_T2_KAS5U
│  ├── mutect2
│  │  └── MMRF_1499_2_PB_Whole_C7_KHS5U-MMRF_1499_1_BM_CD138pos_T2_KAS5U
│  ├── octopus
│  │  └── MMRF_1499_2_PB_Whole_C7_KHS5U-MMRF_1499_1_BM_CD138pos_T2_KAS5U
│  ├── strelka2
│  │  └── MMRF_1499_2_PB_Whole_C7_KHS5U-MMRF_1499_1_BM_CD138pos_T2_KAS5U
│  ├── vardict
│  │  └── MMRF_1499_2_PB_Whole_C7_KHS5U-MMRF_1499_1_BM_CD138pos_T2_KAS5U
│  └── vcfmerger2
│  └── MMRF_1499_2_PB_Whole_C7_KHS5U-MMRF_1499_1_BM_CD138pos_T2_KAS5U
├── genome
│  ├── alignment
│  │  └── bwa
│  │  ├── MMRF_1499_1_BM_CD138pos_T2_KAWGL
│  │  │  └── stats
│  │  └── MMRF_1499_2_PB_Whole_C1_KAWGL
│  │  └── stats
│  ├── constitutional_structural_calls
│  │  └── manta
│  │  └── MMRF_1499_2_PB_Whole_C1_KAWGL
│  ├── constitutional_variant_calls
│  │  ├── deepvariant
│  │  │  └── MMRF_1499_2_PB_Whole_C1_KAWGL
│  │  └── haplotypecaller
│  │  └── MMRF_1499_2_PB_Whole_C1_KAWGL
│  ├── copy_number_analysis
│  │  └── ichorCNA
│  │  ├── MMRF_1499_1_BM_CD138pos_T2_KAWGL
│  │  └── MMRF_1499_2_PB_Whole_C1_KAWGL
│  ├── somatic_copy_number
│  │  └── gatk
│  │  └── MMRF_1499_2_PB_Whole_C1_KAWGL-MMRF_1499_1_BM_CD138pos_T2_KAWGL
│  ├── somatic_structural_calls
│  │  ├── delly
│  │  │  └── MMRF_1499_2_PB_Whole_C1_KAWGL-MMRF_1499_1_BM_CD138pos_T2_KAWGL
│  │  ├── manta
│  │  │  └── MMRF_1499_2_PB_Whole_C1_KAWGL-MMRF_1499_1_BM_CD138pos_T2_KAWGL
│  │  └── pairoscope
│  │  └── MMRF_1499_1_BM_CD138pos_T2_KAWGL
│  └── somatic_variant_calls
│  ├── mutect2
│  │  └── MMRF_1499_2_PB_Whole_C1_KAWGL-MMRF_1499_1_BM_CD138pos_T2_KAWGL
│  ├── octopus
│  │  └── MMRF_1499_2_PB_Whole_C1_KAWGL-MMRF_1499_1_BM_CD138pos_T2_KAWGL
│  ├── strelka2
│  │  └── MMRF_1499_2_PB_Whole_C1_KAWGL-MMRF_1499_1_BM_CD138pos_T2_KAWGL
│  ├── vardict
│  │  └── MMRF_1499_2_PB_Whole_C1_KAWGL-MMRF_1499_1_BM_CD138pos_T2_KAWGL
│  └── vcfmerger2
│  └── MMRF_1499_2_PB_Whole_C1_KAWGL-MMRF_1499_1_BM_CD138pos_T2_KAWGL
├── igv_symbolic_links
├── jetstream
│  ├── history
│  └── logs
├── qc
│  └── multiqc_data
└── rna
├── alignment
│  └── star
│  └── MMRF_1499_1_BM_CD138pos_T1_TSMRU
│  └── stats
├── fusions
│  └── starfusion
│  └── MMRF_1499_1_BM_CD138pos_T1_TSMRU
└── quant
├── salmon
│  └── MMRF_1499_1_BM_CD138pos_T1_TSMRU
└── star
└── MMRF_1499_1_BM_CD138pos_T1_TSMRU
Click to show details
Public Tools Used by the Workflow
All tools build with public easybuild configuration files, available here.
Last Updated July 29th, 2021
| Tool | Version Implemented | Current Version | Dependency and Notes | EasyBuild |
|---|---|---|---|---|
| bcftools | 1.10.2 | 1.13 | Yes | |
| bedtools | 2.29.0 | 2.30.0 | delly-filter, addmatchRNA, vardict, vcfmerger2 | Yes |
| bowtie2 | 2.3.5.1 | 2.4.4 | star-fusion | Yes |
| bwa | 0.7.17 | 0.7.17 | Yes | |
| cellranger | 6.0 | 6.0 | Yes | |
| cuda-toolkit | 10.1.243 | 11.4 | No | |
| deepvariant | 0.10.0 | 1.2.0 | singularity container | Yes |
| delly | 0.7.6 | 0.8.7 | staying with 0.7.6 for compatibility reasons | Yes |
| dle | 0.1.5 | 0.1.5 | private repo | No |
| freebayes | 1.3.1 | 1.3.4 | Yes | |
| gatk | 4.1.8.0 | 4.2.0.0 | Yes | |
| gmap-gsnp | 2019-09-12 | 2021-07-23 | star-fusion | Yes |
| gridss | 2.6.3 | 2.12.0 | ||
| hmmcopyutils | 1.0 | 1.0 | no official release | Yes |
| htseq | 0.12.3 | 0.13.5 | ||
| htslib | 1.10.2 | 1.13 | star-fusion(bgzip) | Yes |
| ichor | 0.2.0 | 0.2.0 | package in R/3.6.1-phoenix module | Yes? |
| jellyfish | 2.3.0 | 2.3.0 | star-fusion | Yes |
| lancet | 1.1.0 | 1.1.0 | Yes | |
| lumosVar2 | 1.1 | 1.1 | Yes | |
| manta | 1.6.0 | 1.6.0 | Yes | |
| manta_tgen | 1.6.0 | 1.6.0 | modified for internal use | No |
| multiQC | 1.9 | 1.11 | python3 pip | Yes |
| octopus | 0.6.3-beta | 0.7.4 | Yes | |
| openmpi | 3.1.3 | 4.1.1 | delly | No |
| pairoscope | 0.4.2 | 0.4.2 | Yes | |
| perl | 5.28.1 | 5.35.2 | star-fusion | Yes |
| phaser | 1.1.1 | 1.1.1 | vcfmerger2 | Yes |
| python2 | 2.7.15 | 2.7.18 | Yes | |
| python3 | 3.7.2 | 3.9.6 | star-fusion, vcfmerger2 | Yes |
| R | 3.6.1 | 4.1.0 | gatk cnv, varDict, vcfmerger2 | Yes |
| salmon | 1.2.1 | 1.5.2 | self, star-fusion | Yes |
| sambamba | 0.7.0 | 0.8.0 | ||
| samblaster | 0.1.24 | 0.1.26 | ||
| samtools | 1.10 | 1.13 | Yes | |
| singularity | 3.5.2 | 3.8.1 | deepvariant | Yes |
| snpEff | 4.3t | 4.5covid19 | covid related release? | Yes |
| snpSniffer | 7.0.0 | 7.0.0 | No | |
| star | 2.7.5a | 2.7.9a | self, star-fusion | Yes |
| star-fusion | 1.8.1 | 1.10.1 | Yes | |
| strelka | 2.9.10 | 2.9.10 | Yes | |
| subread | 2.0.0 | 2.0.3 | part of subread package | Yes |
| tgen_mutation_burden | 1.2.3 | 1.2.3 | No | |
| transParser | 1.0.1 | 1.0.1 | No | |
| trinityrnaseq | 2.8.6 | 2.12.0 | star-fusion | Yes |
| vardictJava | 1.7.0 | 1.8.2 | Yes | |
| vcfmerger2 | 0.8.7 | 0.8.7 | Yes | |
| vep | 98.3 | 104.3 | Yes | |
| verifybamid2 | 1.0.6 | 2.0.1 | Yes | |
| vt | 0_57721 | 0_57721 | Yes |
Required PERL Modules
| Module | Version |
|---|---|
| AutoLoader | 5.74 |
| Bio::EnsEMBL::XS | 2.3.2 |
| Carp::Assert | 0.21 |
| common::sense | 3.74 |
| constant | 1.33 |
| Cwd | 3.75 |
| Data::Dumper | 2.173 |
| Data::Dumper | 2.173 |
| DB_File | 1.852 |
| Encode | 3.01 |
| ExtUtils::CBuilder | 0.280231 |
| ExtUtils::CppGuess | 0.19 |
| ExtUtils::MakeMaker | 7.36 |
| ExtUtils::ParseXS | 3.35 |
| IO::File | 1.39 |
| IO::Select | 1.39 |
| IO::Socket | 1.39 |
| IPC::Cmd | 1.02 |
| JSON::XS | 4.02 |
| List::Util | 1.5 |
| Locale::Maketext::Simple | 0.21 |
| MIME::Base64 | 3.15 |
| Module::CoreList | 5.2019062 |
| Module::Load | 0.34 |
| Module::Load::Conditional | 0.68 |
| Module::Metadata | 1.000036 |
| Net::Domain | 3.11 |
| Params::Check | 0.38 |
| parent | 0.237 |
| Perl::OSType | 1.01 |
| PerlIO::gzip | 0.2 |
| Pod::Escapes | 1.07 |
| Pod::Man | 4.12 |
| Pod::Simple | 3.39 |
| Scalar::Util | 1.5 |
| Set::IntervalTree | 0.12 |
| Socket | 2.029 |
| Storable | 3.15 |
| Test | 1.26 |
| Test::More | 1.302164 |
| Text::ParseWords | 3.3 |
| Text::Wrap | 2013.0523 |
| Time::Local | 1.28 |
| Types::Serialiser | 1 |
| URI::Escape | 1.76 |
| XSLoader | 0.24 |
Required PYTHON2 Modules
| Module | Version |
|---|---|
| alabaster | 0.7.12 |
| asn1crypto | 0.24.0 |
| atomicwrites | 1.3.0 |
| attrs | 19.1.0 |
| Babel | 2.6.0 |
| bcrypt | 3.1.6 |
| bitstring | 3.1.5 |
| blist | 1.3.6 |
| certifi | 2019.3.9 |
| cffi | 1.12.2 |
| chardet | 3.0.4 |
| Click | 7 |
| cryptography | 2.6.1 |
| Cython | 0.29.6 |
| deap | 1.2.2 |
| decorator | 4.3.2 |
| docopt | 0.6.2 |
| docutils | 0.14 |
| ecdsa | 0.13 |
| enum34 | 1.1.6 |
| funcsigs | 1.0.2 |
| future | 0.17.1 |
| idna | 2.8 |
| imagesize | 1.1.0 |
| ipaddress | 1.0.22 |
| Jinja2 | 2.1 |
| joblib | 0.13.2 |
| liac-arff | 2.4.0 |
| MarkupSafe | 1.1.1 |
| mock | 2.0.0 |
| more-itertools | 5.0.0 |
| netaddr | 0.7.19 |
| netifaces | 0.10.9 |
| nose | 1.3.7 |
| packaging | 19 |
| paramiko | 2.4.2 |
| pathlib2 | 2.3.3 |
| paycheck | 1.0.2 |
| pbr | 5.1.3 |
| pip | 19.0.3 |
| pluggy | 0.9.0 |
| psutil | 5.6.1 |
| py | 1.8.0 |
| py_expression_eval | 0.3.6 |
| pyasn1 | 0.4.5 |
| pycparser | 2.19 |
| pycrypto | 2.6.1 |
| Pygments | 2.3.1 |
| PyNaCl | 1.3.0 |
| pyparsing | 2.3.1 |
| pytest | 4.3.1 |
| python-dateutil | 2.8.0 |
| pytz | 2018.9 |
| requests | 2.21.0 |
| scandir | 1.10.0 |
| setuptools | 40.8.0 |
| setuptools_scm | 3.2.0 |
| singledispatch | 3.4.0.3 |
| six | 1.12.0 |
| snowballstemmer | 1.2.1 |
| Sphinx | 1.8.5 |
| sphinxcontrib-websupport | 1.1.0 |
| tabulate | 0.8.3 |
| typing | 3.6.6 |
| ujson | 1.35 |
| urllib3 | 1.24.1 |
| virtualenv | 16.4.3 |
| wheel | 0.33.1 |
| xlrd | 1.2.0 |
Required PYTHON3 Modules
- cyvcf2
- getopt
- igv_reports
- multiprocessing
- multiqc
- PIL
- pybedtools
- pysam
- requests
- sys (argv, exit)
Required R Libraries
- ichorCNA2.0
- tidyverse (ggplot)
Please see the wiki for a detailed install guide
In order to run from the command line, we need to create a config file for our project. The general format is as follows:
{
"project": "",
"study": "",
"email": "",
"hpcAccount": "",
"isilonPath": "",
"pipeline": "coyote",
"dataFiles": [],
"dnaAlignmentStyle": "tgen",
"email": "[email protected]",
"isilonPath": "",
"pipeline": "coyote@version",
"project": "Project_Name",
"submitter": "somebody",
"tasks": {},
}Here is a larger example with actual data for running the coyote pipeline on a NA12878 project:
NA12878 Example
{
"cram": true,
"dataFiles": [
{
"assayCode": "TPFWG",
"dnaRnaMergeKey": "GIAB_NA12878_1_CL_Whole",
"fastqCode": "R1",
"fastqPath": "/home/tgenref/homo_sapiens/control_files/giab/fastq/NA12878_140407_D00360_0016_ASUPERFQS01/Project_GIAB_NA12878_1_TPFWG/Sample_GIAB_NA12878_1_CL_Whole_C1_TPFWG_K18088_SUPERFQS01/GIAB_NA12878_1_CL_Whole_C1_TPFWG_K18088_SUPERFQS01_NoIndex_L001_R1_001.fastq.gz",
"fileType": "fastq",
"fraction": "Whole",
"glPrep": "Genome",
"glType": "Genome",
"index1Length": 6,
"index2Length": 0,
"limsLibraryRecordId": 64391,
"numberOfReads": 228228468,
"read1Length": 148,
"read2Length": 148,
"readOrientation": "Inward",
"rgcn": "TGen",
"rgid": "SUPERFQS01_1_K18088",
"rgbc": "ATCACG",
"rglb": "K18088",
"rgpl": "ILLUMINA",
"rgpm": "HiSeq2500",
"rgpu": "SUPERFQS01_1",
"rgsm": "GIAB_NA12878_1_CL_Whole_C1",
"rnaStrandDirection": "NotApplicable",
"rnaStrandType": "NotApplicable",
"sampleMergeKey": "GIAB_NA12878_1_CL_Whole_C1_TPFWG",
"sampleName": "GIAB_NA12878_1_CL_Whole_C1_TPFWG_K18088",
"subGroup": "Constitutional",
"umiInLine": "false",
"umiLength": 0,
"umiRead": false
},
{
"assayCode": "TPFWG",
"dnaRnaMergeKey": "GIAB_NA12878_1_CL_Whole",
"fastqCode": "R2",
"fastqPath": "/home/tgenref/homo_sapiens/control_files/giab/fastq/NA12878_140407_D00360_0016_ASUPERFQS01/Project_GIAB_NA12878_1_TPFWG/Sample_GIAB_NA12878_1_CL_Whole_C1_TPFWG_K18088_SUPERFQS01/GIAB_NA12878_1_CL_Whole_C1_TPFWG_K18088_SUPERFQS01_NoIndex_L001_R2_001.fastq.gz",
"fileType": "fastq",
"fraction": "Whole",
"glPrep": "Genome",
"glType": "Genome",
"index1Length": 6,
"index2Length": 0,
"limsLibraryRecordId": 64391,
"numberOfReads": 228228468,
"read1Length": 148,
"read2Length": 148,
"readOrientation": "Inward",
"rgcn": "TGen",
"rgid": "SUPERFQS01_1_K18088",
"rgbc": "ATCACG",
"rglb": "K18088",
"rgpl": "ILLUMINA",
"rgpm": "HiSeq2500",
"rgpu": "SUPERFQS01_1",
"rgsm": "GIAB_NA12878_1_CL_Whole_C1",
"rnaStrandDirection": "NotApplicable",
"rnaStrandType": "NotApplicable",
"sampleMergeKey": "GIAB_NA12878_1_CL_Whole_C1_TPFWG",
"sampleName": "GIAB_NA12878_1_CL_Whole_C1_TPFWG_K18088",
"subGroup": "Constitutional",
"umiInLine": "false",
"umiLength": 0,
"umiRead": false
},
{
"assayCode": "TPFWG",
"dnaRnaMergeKey": "GIAB_NA12878_1_CL_Whole",
"fastqCode": "R1",
"fastqPath": "/home/tgenref/homo_sapiens/control_files/giab/fastq/NA12878_140407_D00360_0016_ASUPERFQS01/Project_GIAB_NA12878_1_TPFWG/Sample_GIAB_NA12878_1_CL_Whole_C1_TPFWG_K18088_SUPERFQS01/GIAB_NA12878_1_CL_Whole_C1_TPFWG_K18088_SUPERFQS01_NoIndex_L002_R1_001.fastq.gz",
"fileType": "fastq",
"fraction": "Whole",
"glPrep": "Genome",
"glType": "Genome",
"index1Length": 6,
"index2Length": 0,
"limsLibraryRecordId": 64391,
"numberOfReads": 232329584,
"read1Length": 148,
"read2Length": 148,
"readOrientation": "Inward",
"rgcn": "TGen",
"rgid": "SUPERFQS01_2_K18088",
"rgbc": "ATCACG",
"rglb": "K18088",
"rgpl": "ILLUMINA",
"rgpm": "HiSeq2500",
"rgpu": "SUPERFQS01_2",
"rgsm": "GIAB_NA12878_1_CL_Whole_C1",
"rnaStrandDirection": "NotApplicable",
"rnaStrandType": "NotApplicable",
"sampleMergeKey": "GIAB_NA12878_1_CL_Whole_C1_TPFWG",
"sampleName": "GIAB_NA12878_1_CL_Whole_C1_TPFWG_K18088",
"subGroup": "Constitutional",
"umiInLine": "false",
"umiLength": 0,
"umiRead": false
},
{
"assayCode": "TPFWG",
"dnaRnaMergeKey": "GIAB_NA12878_1_CL_Whole",
"fastqCode": "R2",
"fastqPath": "/home/tgenref/homo_sapiens/control_files/giab/fastq/NA12878_140407_D00360_0016_ASUPERFQS01/Project_GIAB_NA12878_1_TPFWG/Sample_GIAB_NA12878_1_CL_Whole_C1_TPFWG_K18088_SUPERFQS01/GIAB_NA12878_1_CL_Whole_C1_TPFWG_K18088_SUPERFQS01_NoIndex_L002_R2_001.fastq.gz",
"fileType": "fastq",
"fraction": "Whole",
"glPrep": "Genome",
"glType": "Genome",
"index1Length": 6,
"index2Length": 0,
"limsLibraryRecordId": 64391,
"numberOfReads": 232329584,
"read1Length": 148,
"read2Length": 148,
"readOrientation": "Inward",
"rgcn": "TGen",
"rgid": "SUPERFQS01_2_K18088",
"rgbc": "ATCACG",
"rglb": "K18088",
"rgpl": "ILLUMINA",
"rgpm": "HiSeq2500",
"rgpu": "SUPERFQS01_2",
"rgsm": "GIAB_NA12878_1_CL_Whole_C1",
"rnaStrandDirection": "NotApplicable",
"rnaStrandType": "NotApplicable",
"sampleMergeKey": "GIAB_NA12878_1_CL_Whole_C1_TPFWG",
"sampleName": "GIAB_NA12878_1_CL_Whole_C1_TPFWG_K18088",
"subGroup": "Constitutional",
"umiInLine": "false",
"umiLength": 0,
"umiRead": false
},
{
"assayCode": "TPFWG",
"dnaRnaMergeKey": "GIAB_NA12878_1_CL_Whole",
"fastqCode": "R1",
"fastqPath": "/home/tgenref/homo_sapiens/control_files/giab/fastq/NA12878_140407_D00360_0017_BSUPERFQS02/Project_GIAB_NA12878_1_TPFWG/Sample_GIAB_NA12878_1_CL_Whole_C1_TPFWG_K18088_SUPERFQS02/GIAB_NA12878_1_CL_Whole_C1_TPFWG_K18088_SUPERFQS02_NoIndex_L001_R1_001.fastq.gz",
"fileType": "fastq",
"fraction": "Whole",
"glPrep": "Genome",
"glType": "Genome",
"index1Length": 6,
"index2Length": 0,
"limsLibraryRecordId": 64391,
"numberOfReads": 231481708,
"read1Length": 148,
"read2Length": 148,
"readOrientation": "Inward",
"rgcn": "TGen",
"rgid": "SUPERFQS02_1_K18088",
"rgbc": "ATCACG",
"rglb": "K18088",
"rgpl": "ILLUMINA",
"rgpm": "HiSeq2500",
"rgpu": "SUPERFQS02_1",
"rgsm": "GIAB_NA12878_1_CL_Whole_C1",
"rnaStrandDirection": "NotApplicable",
"rnaStrandType": "NotApplicable",
"sampleMergeKey": "GIAB_NA12878_1_CL_Whole_C1_TPFWG",
"sampleName": "GIAB_NA12878_1_CL_Whole_C1_TPFWG_K18088",
"subGroup": "Constitutional",
"umiInLine": "false",
"umiLength": 0,
"umiRead": false
},
{
"assayCode": "TPFWG",
"dnaRnaMergeKey": "GIAB_NA12878_1_CL_Whole",
"fastqCode": "R2",
"fastqPath": "/home/tgenref/homo_sapiens/control_files/giab/fastq/NA12878_140407_D00360_0017_BSUPERFQS02/Project_GIAB_NA12878_1_TPFWG/Sample_GIAB_NA12878_1_CL_Whole_C1_TPFWG_K18088_SUPERFQS02/GIAB_NA12878_1_CL_Whole_C1_TPFWG_K18088_SUPERFQS02_NoIndex_L001_R2_001.fastq.gz",
"fileType": "fastq",
"fraction": "Whole",
"glPrep": "Genome",
"glType": "Genome",
"index1Length": 6,
"index2Length": 0,
"limsLibraryRecordId": 64391,
"numberOfReads": 231481708,
"read1Length": 148,
"read2Length": 148,
"readOrientation": "Inward",
"rgcn": "TGen",
"rgid": "SUPERFQS02_1_K18088",
"rgbc": "ATCACG",
"rglb": "K18088",
"rgpl": "ILLUMINA",
"rgpm": "HiSeq2500",
"rgpu": "SUPERFQS02_1",
"rgsm": "GIAB_NA12878_1_CL_Whole_C1",
"rnaStrandDirection": "NotApplicable",
"rnaStrandType": "NotApplicable",
"sampleMergeKey": "GIAB_NA12878_1_CL_Whole_C1_TPFWG",
"sampleName": "GIAB_NA12878_1_CL_Whole_C1_TPFWG_K18088",
"subGroup": "Constitutional",
"umiInLine": "false",
"umiLength": 0,
"umiRead": false
},
{
"assayCode": "TPFWG",
"dnaRnaMergeKey": "GIAB_NA12878_1_CL_Whole",
"fastqCode": "R1",
"fastqPath": "/home/tgenref/homo_sapiens/control_files/giab/fastq/NA12878_140407_D00360_0017_BSUPERFQS02/Project_GIAB_NA12878_1_TPFWG/Sample_GIAB_NA12878_1_CL_Whole_C1_TPFWG_K18088_SUPERFQS02/GIAB_NA12878_1_CL_Whole_C1_TPFWG_K18088_SUPERFQS02_NoIndex_L002_R1_001.fastq.gz",
"fileType": "fastq",
"fraction": "Whole",
"glPrep": "Genome",
"glType": "Genome",
"index1Length": 6,
"index2Length": 0,
"limsLibraryRecordId": 64391,
"numberOfReads": 228988006,
"read1Length": 148,
"read2Length": 148,
"readOrientation": "Inward",
"rgcn": "TGen",
"rgid": "SUPERFQS02_2_K18088",
"rgbc": "ATCACG",
"rglb": "K18088",
"rgpl": "ILLUMINA",
"rgpm": "HiSeq2500",
"rgpu": "SUPERFQS02_2",
"rgsm": "GIAB_NA12878_1_CL_Whole_C1",
"rnaStrandDirection": "NotApplicable",
"rnaStrandType": "NotApplicable",
"sampleMergeKey": "GIAB_NA12878_1_CL_Whole_C1_TPFWG",
"sampleName": "GIAB_NA12878_1_CL_Whole_C1_TPFWG_K18088",
"subGroup": "Constitutional",
"umiInLine": "false",
"umiLength": 0,
"umiRead": false
},
{
"assayCode": "TPFWG",
"dnaRnaMergeKey": "GIAB_NA12878_1_CL_Whole",
"fastqCode": "R2",
"fastqPath": "/home/tgenref/homo_sapiens/control_files/giab/fastq/NA12878_140407_D00360_0017_BSUPERFQS02/Project_GIAB_NA12878_1_TPFWG/Sample_GIAB_NA12878_1_CL_Whole_C1_TPFWG_K18088_SUPERFQS02/GIAB_NA12878_1_CL_Whole_C1_TPFWG_K18088_SUPERFQS02_NoIndex_L002_R2_001.fastq.gz",
"fileType": "fastq",
"fraction": "Whole",
"glPrep": "Genome",
"glType": "Genome",
"index1Length": 6,
"index2Length": 0,
"limsLibraryRecordId": 64391,
"numberOfReads": 228988006,
"read1Length": 148,
"read2Length": 148,
"readOrientation": "Inward",
"rgcn": "TGen",
"rgid": "SUPERFQS02_2_K18088",
"rgbc": "ATCACG",
"rglb": "K18088",
"rgpl": "ILLUMINA",
"rgpm": "HiSeq2500",
"rgpu": "SUPERFQS02_2",
"rgsm": "GIAB_NA12878_1_CL_Whole_C1",
"rnaStrandDirection": "NotApplicable",
"rnaStrandType": "NotApplicable",
"sampleMergeKey": "GIAB_NA12878_1_CL_Whole_C1_TPFWG",
"sampleName": "GIAB_NA12878_1_CL_Whole_C1_TPFWG_K18088",
"subGroup": "Constitutional",
"umiInLine": "false",
"umiLength": 0,
"umiRead": false
}
],
"dnaAlignmentStyle": "tgen",
"email": "[email protected]",
"holdConfig": false,
"hpcAccount": "tgen-#####",
"isilonPath": "/example/giab/",
"matchedNormal": true,
"matchedNormalToUse": "",
"maternalID": "",
"patCode": "NA12878",
"paternalID": "",
"pipeline": "coyote",
"project": "GIAB_NA12878",
"sex": "Female",
"study": "GIAB",
"submitter": "user",
"submitterEmail": "[email protected]",
"tasks": {
"Exome_alignment_base_recalibration_gatk" : false,
"Exome_alignment_mark_duplicates_gatk" : false,
"Exome_alignment_mark_duplicates_samblaster" : false,
"Exome_alignment_mark_duplicates_samtools" : true,
"Exome_constitutional_annotate_vcfs_bcftools_eva_gca" : true,
"Exome_constitutional_annotate_vcfs_snpEff_ann" : true,
"Exome_constitutional_annotate_vcfs_vep" : true,
"Exome_constitutional_genotype_hc_gvcf_gatk_GenotypeGVCFs" : true,
"Exome_constitutional_snp_indel_caller_deepvariant" : true,
"Exome_constitutional_snp_indel_caller_freebayes" : true,
"Exome_constitutional_snp_indel_caller_gatk_HaplotypeCaller" : true,
"Exome_constitutional_snp_indel_caller_octopus" : true,
"Exome_constitutional_snp_indel_caller_strelka2" : true,
"Exome_constitutional_structural_caller_manta" : true,
"Exome_quality_control_constitutional_contamination_check_VerifyBamID" : true,
"Exome_quality_control_constitutional_sex_check_freebayes" : true,
"Exome_quality_control_genotype_concordance_snpSniffer" : true,
"Exome_quality_control_stats_gatk_CollectHsMetrics" : true,
"Exome_quality_control_stats_gatk_CollectMultipleMetrics" : true,
"Exome_quality_control_stats_gatk_ConvertSequencingArtifactToOxoG" : true,
"Exome_quality_control_stats_samtools_flagstat" : false,
"Exome_quality_control_stats_samtools_idxstats" : true,
"Exome_quality_control_stats_samtools_markdup_stats" : true,
"Exome_quality_control_stats_samtools_stats" : true,
"Exome_somatic_annotate_vcfs_bcftools_clinvar": true,
"Exome_somatic_annotate_vcfs_bcftools_cosmic_coding": true,
"Exome_somatic_annotate_vcfs_bcftools_cosmic_noncoding": true,
"Exome_somatic_annotate_vcfs_bcftools_dbsnp": true,
"Exome_somatic_annotate_vcfs_bcftools_gnomad_exome": true,
"Exome_somatic_annotate_vcfs_bcftools_gnomad_genome": true,
"Exome_somatic_annotate_vcfs_bcftools_topmed": true,
"Exome_somatic_annotate_vcfs_snpEff_ann": true,
"Exome_somatic_annotate_vcfs_vep": true,
"Exome_somatic_cna_caller_gatk": true,
"Exome_somatic_merge_vcfs_vcfMerger2": true,
"Exome_somatic_sample_metric_msisensor_pro": true,
"Exome_somatic_sample_metric_tgen_mutation_burden": true,
"Exome_somatic_snp_indel_caller_VarDict": true,
"Exome_somatic_snp_indel_caller_gatk_mutect2": true,
"Exome_somatic_snp_indel_caller_lancet": true,
"Exome_somatic_snp_indel_caller_octopus": true,
"Exome_somatic_snp_indel_caller_strelka2": true,
"Exome_somatic_structural_caller_delly": false,
"Exome_somatic_structural_caller_gridss": false,
"Exome_somatic_structural_caller_manta": true,
"Exome_tumor_only_annotate_vcfs_bcftools_clinvar": true,
"Exome_tumor_only_annotate_vcfs_bcftools_cosmic_coding": true,
"Exome_tumor_only_annotate_vcfs_bcftools_cosmic_noncoding": true,
"Exome_tumor_only_annotate_vcfs_bcftools_dbsnp": true,
"Exome_tumor_only_annotate_vcfs_bcftools_gnomad_exome": true,
"Exome_tumor_only_annotate_vcfs_bcftools_gnomad_genome": true,
"Exome_tumor_only_annotate_vcfs_bcftools_topmed": true,
"Exome_tumor_only_annotate_vcfs_snpEff_ann": true,
"Exome_tumor_only_annotate_vcfs_vep": true,
"Exome_tumor_only_cna_caller_gatk": true,
"Exome_tumor_only_merge_vcfs_vcfMerger2": true,
"Exome_tumor_only_snp_indel_caller_VarDict": true,
"Exome_tumor_only_snp_indel_caller_deepvariant": true,
"Exome_tumor_only_snp_indel_caller_gatk_mutect2": true,
"Exome_tumor_only_snp_indel_caller_lancet": true,
"Exome_tumor_only_snp_indel_caller_lumosvar2": false,
"Exome_tumor_only_snp_indel_caller_octopus": true,
"Exome_tumor_only_snp_indel_caller_strelka2": true,
"Exome_tumor_only_structural_caller_delly": false,
"Exome_tumor_only_structural_caller_gridss": false,
"Exome_tumor_only_structural_caller_manta": true,
"Genome_alignment_base_recalibration_gatk" : false,
"Genome_alignment_mark_duplicates_gatk" : false,
"Genome_alignment_mark_duplicates_samblaster" : false,
"Genome_alignment_mark_duplicates_samtools" : true,
"Genome_constitutional_annotate_vcfs_bcftools_eva_gca" : true,
"Genome_constitutional_annotate_vcfs_snpEff_ann" : true,
"Genome_constitutional_annotate_vcfs_vep" : true,
"Genome_constitutional_cna_caller_ichor" : true,
"Genome_constitutional_genotype_hc_gvcf_gatk_GenotypeGVCFs" : true,
"Genome_constitutional_snp_indel_caller_deepvariant" : true,
"Genome_constitutional_snp_indel_caller_freebayes" : true,
"Genome_constitutional_snp_indel_caller_gatk_HaplotypeCaller" : true,
"Genome_constitutional_snp_indel_caller_octopus" : true,
"Genome_constitutional_snp_indel_caller_strelka2" : true,
"Genome_constitutional_structural_caller_manta" : true,
"Genome_quality_control_constitutional_contamination_check_VerifyBamID" : true,
"Genome_quality_control_constitutional_sex_check_freebayes" : true,
"Genome_quality_control_genotype_concordance_snpSniffer" : true,
"Genome_quality_control_stats_gatk_CollectMultipleMetrics" : true,
"Genome_quality_control_stats_gatk_CollectRawWgsMetrics" : true,
"Genome_quality_control_stats_gatk_CollectWgsMetrics" : true,
"Genome_quality_control_stats_gatk_CollectWgsMetricsWithNonZeroCoverage" : true,
"Genome_quality_control_stats_gatk_ConvertSequencingArtifactToOxoG" : true,
"Genome_quality_control_stats_samtools_flagstat" : true,
"Genome_quality_control_stats_samtools_idxstats" : true,
"Genome_quality_control_stats_samtools_stats" : true,
"Genome_somatic_annotate_vcfs_bcftools_eva_gca" : true,
"Genome_somatic_annotate_vcfs_snpEff_ann" : true,
"Genome_somatic_annotate_vcfs_vep" : true,
"Genome_somatic_cna_caller_gatk" : true,
"Genome_somatic_merge_vcfs_vcfMerger2" : true,
"Genome_somatic_sample_metric_msisensor_pro" : true,
"Genome_somatic_sample_metric_tgen_mutation_burden" : true,
"Genome_somatic_snp_indel_caller_VarDict" : true,
"Genome_somatic_snp_indel_caller_gatk_mutect2" : true,
"Genome_somatic_snp_indel_caller_lancet" : false,
"Genome_somatic_snp_indel_caller_octopus" : true,
"Genome_somatic_snp_indel_caller_strelka2" : true,
"Genome_somatic_structural_caller_delly" : false,
"Genome_somatic_structural_caller_gridss" : false,
"Genome_somatic_structural_caller_manta" : true,
"Genome_tumor_only_annotate_vcfs_bcftools_clinvar" : true,
"Genome_tumor_only_annotate_vcfs_bcftools_cosmic_coding" : true,
"Genome_tumor_only_annotate_vcfs_bcftools_cosmic_noncoding" : true,
"Genome_tumor_only_annotate_vcfs_bcftools_dbsnp" : true,
"Genome_tumor_only_annotate_vcfs_bcftools_gnomad_exome" : true,
"Genome_tumor_only_annotate_vcfs_bcftools_gnomad_genome" : true,
"Genome_tumor_only_annotate_vcfs_bcftools_topmed" : true,
"Genome_tumor_only_annotate_vcfs_snpEff_ann" : true,
"Genome_tumor_only_annotate_vcfs_vep" : true,
"Genome_tumor_only_cna_caller_gatk" : true,
"Genome_tumor_only_merge_vcfs_vcfMerger2" : false,
"Genome_tumor_only_snp_indel_caller_VarDict" : false,
"Genome_tumor_only_snp_indel_caller_deepvariant" : true,
"Genome_tumor_only_snp_indel_caller_gatk_mutect2" : false,
"Genome_tumor_only_snp_indel_caller_lancet" : false,
"Genome_tumor_only_snp_indel_caller_octopus" : false,
"Genome_tumor_only_snp_indel_caller_strelka2" : false,
"Genome_tumor_only_structural_caller_delly" : false,
"Genome_tumor_only_structural_caller_gridss" : false,
"Genome_tumor_only_structural_caller_manta" : true,
"RNA_alignment_rna_alignment_STAR" : true,
"RNA_quality_control_genotype_concordance_snpSniffer" : true,
"RNA_quality_control_stats_gatk_CollectMultipleMetrics" : true,
"RNA_quality_control_stats_gatk_CollectRnaSeqMetrics" : true,
"RNA_quality_control_stats_gatk_ConvertSequencingArtifactToOxoG" : false,
"RNA_quality_control_stats_samtools_flagstat" : false,
"RNA_quality_control_stats_samtools_idxstats" : true,
"RNA_quality_control_stats_samtools_stats" : true,
"RNA_sample_metric_sample_metric_samtools_bedcov" : true,
"RNA_transcriptome_fusion_caller_Keep_STAR_Fusion_BAM" : false,
"RNA_transcriptome_fusion_caller_STAR_Fusion" : true,
"RNA_transcriptome_quantify_expression_HTseq" : true,
"RNA_transcriptome_quantify_expression_RSEM" : false,
"RNA_transcriptome_quantify_expression_salmon_bam" : false,
"RNA_transcriptome_quantify_expression_salmon_fastqs" : true,
"RNA_transcriptome_quantify_expression_subread_featureCounts" : false,
"SingleCellRNA_VDJ_Assembly_cellranger_vdj" : true,
"SingleCellRNA_quality_control_genotype_concordance_snpSniffer" : true,
"SingleCellRNA_quality_control_stats_gatk_CollectMultipleMetrics" : true,
"SingleCellRNA_quality_control_stats_gatk_CollectRnaSeqMetrics" : true,
"SingleCellRNA_quality_control_stats_gatk_ConvertSequencingArtifactToOxoG" : true,
"SingleCellRNA_quality_control_stats_samtools_flagstat" : true,
"SingleCellRNA_quality_control_stats_samtools_idxstats" : true,
"SingleCellRNA_quality_control_stats_samtools_stats" : true,
"SingleCellRNA_sample_metric_sample_metric_samtools_bedcov" : true,
"SingleCellRNA_transcriptome_quantify_expression_STARsolo" : true,
"SingleCellRNA_transcriptome_quantify_expression_cellranger_count" : true
},
"varDB": false
}Looking at the block of objects between the datafiles and the tasks one might notice some objects not mentioned in the minimal example provided in the general example. Some of these might be specific to the project and your environment. The common ones that we use in our primary use case are:
"dnaAlignmentStyle": "",
"email": "",
"ethnicity": "",
"familyCode": "",
"holdConfig": false,
"hpcAccount": "",
"isilonPath": "",
"matchedNormal": true,
"matchedNormalToUse": "",
"maternalID": "",
"patCode": "",
"paternalID": "",
"pipeline": "",
"project": "",
"sex": "",
"study": "",
"submissionSource": "",
"submitter": "",Once we have a config file for the project we're ready to initialize and launch the project. We can initialize a project via
$ jetstream init -h
usage: jetstream init [-h] [-l] [-p PROJECT] [-f] [--project-id PROJECT_ID]
[-c TYPE:KEY VALUE] [-C PATH]
[path]
Create or reinitialize a project This command is used to create a new
Jetstream project directory. If no path is given, the current directory will
be initialized. If config data options are given (-c/--config/--config-file),
they will be added to the project config file.
positional arguments:
path Path to a initialize a project
optional arguments:
-h, --help show this help message and exit
-l , --logging set the logging profile
-p PROJECT, --project PROJECT
path to a Jetstream project directory
-f, --force Force overwrite of project.yaml
--project-id PROJECT_ID
Force a project ID instead of using letting it be
generated automatically
template variables:
These options are used to add data that is available for rendering
templates. These arguments should follow the syntax "-c <[type:]key>
<value>". They can be used multiple times.
-c TYPE:KEY VALUE, --config TYPE:KEY VALUE
add a single template variable
-C PATH, --config-file PATH
load template variables from a file
$ jetstream init GIAB -C GIAB_NA12878_24582bb3f7.json
This creates a jetstream project with the title of GIAB. Now in order to run the coyote pipeline on this project, we need to use:
$ jetstream pipelines -h
usage: jetstream pipelines [-h] [-l] [-p PROJECT] [-o OUT] [-b] [-r]
[--backend {local,slurm}]
[--format {template,module,workflow}]
[--reset-method {retry,resume,reset}]
[--existing-workflow EXISTING_WORKFLOW]
[--template-dir [SEARCH_PATH]] [-c TYPE:KEY VALUE]
[-C PATH] [--pipelines-home PIPELINES_HOME] [-L]
[path]
Run a pipeline. Pipelines are Jetstream templates that have been documented
with version information and added to the jetstream pipelines directory. This
command allows pipelines to be referenced by name and automatically includes
the pipeline scripts and constants in the run. Run Jetstream from a template,
module, or workflow
positional arguments:
path path to a template, module, or workflow file. (if
using "pipelines" command, the name of the pipeline)
optional arguments:
-h, --help show this help message and exit
-l , --logging set the logging profile
-p PROJECT, --project PROJECT
path to a Jetstream project directory
-o OUT, --out OUT path to save the workflow progress (this will be set
automatically if working with a project) [None]
-b, --build-only just render the template, build the workflow, and stop
-r, --render-only just render the template and stop
--backend {local,slurm}
runner backend name used for executing tasks [slurm]
--format {template,module,workflow}
workflow format - if this is None, it will be inferred
from the extension of the path [None]
--reset-method {retry,resume,reset}
controls which tasks are reset prior to starting the
run - "retry": pending and failed, "resume": pending,
or "reset": all [retry]
--existing-workflow EXISTING_WORKFLOW
path to an existing workflow file that will be merged
into run (this will be set automatically if working
with a project)
--template-dir [SEARCH_PATH]
directory to add to search path for loading templates,
this can be used multiple times
template variables:
These options are used to add data that is available for rendering
templates. These arguments should follow the syntax "-c <[type:]key>
<value>". They can be used multiple times.
-c TYPE:KEY VALUE, --config TYPE:KEY VALUE
add a single template variable
-C PATH, --config-file PATH
load template variables from a file
pipeline options:
--pipelines-home PIPELINES_HOME
override path to the pipelines home
-L, --list show a list of all the pipelines installed
$ jetstream pipelines coyote -p GIAB
Now we wait for the pipeline to finish!
For each of our data files/fastqs we have some required data, many of which are self explained, but we will explain the more unique variables. Here is an example:
"dataFiles": [
{
"assayCode": "TPFWG",
"dnaRnaMergeKey": "GIAB_NA12878_1_CL_Whole",
"fastqCode": "R1",
"fastqPath": "/home/tgenref/homo_sapiens/control_files/giab/fastq/NA12878_140407_D00360_0016_ASUPERFQS01/Project_GIAB_NA12878_1_TPFWG/Sample_GIAB_NA12878_1_CL_Whole_C1_TPFWG_K18088_SUPERFQS01/GIAB_NA12878_1_CL_Whole_C1_TPFWG_K18088_SUPERFQS01_NoIndex_L001_R1_001.fastq.gz",
"fileType": "fastq",
"fraction": "Whole",
"glPrep": "Genome",
"glType": "Genome",
"index1Length": 6,
"index2Length": 0,
"limsLibraryRecordId": 64391,
"numberOfReads": 228228468,
"read1Length": 148,
"read2Length": 148,
"readOrientation": "Inward",
"rgcn": "TGen",
"rgid": "SUPERFQS01_1_K18088",
"rgbc": "ATCACG",
"rglb": "K18088",
"rgpl": "ILLUMINA",
"rgpm": "HiSeq2500",
"rgpu": "SUPERFQS01_1",
"rgsm": "GIAB_NA12878_1_CL_Whole_C1",
"rnaStrandDirection": "NotApplicable",
"rnaStrandType": "NotApplicable",
"sampleMergeKey": "GIAB_NA12878_1_CL_Whole_C1_TPFWG",
"sampleName": "GIAB_NA12878_1_CL_Whole_C1_TPFWG_K18088",
"subGroup": "Constitutional",
"umiInLine": "false",
"umiLength": 0,
"umiRead": false
}There are restrictions on what some of these variables can be assigned to, these will be denoted in the [ ]'s. If the attribute isn't strictly required then it is not included in this list.
-
assayCode
Genome: [*] We are not concerned about the assayCode for genomes.
Note: We have a number of bed files supporting our exome captures, these are the shortened capture codes
Exome: [ *SC2 | *XT2 ]
Used for determining if the sample is DNA/RNA/etc. and adding the corresponding tasks to the final workflow. Each sample discovered will take this attribute from the first file encountered for that sample in the config file. -
dnaRnaMergeKey
Used during DNA/RNA integrations steps. It defines the pairing of DNA and RNA samples as a project might have multiple DNA and RNA pairs, for instance it can be used to ensure the diagnosis exome and RNA are paired together and the relapse exome is not paired with the diagnosis RNA. -
fastqCode [R1|R2]
Assigns the read number of the fastq following standard Illumina paired-end nomenclature. -
fastqPath
Assigns the path to the fastq. -
fileType
Assigns the file type. -
glPrep [genome|capture|rna]
Used for determining the prep used to create the sample and then modifying how the pipeline runs depending on the prep. This is used to configure single cell as well as CHIP preps. -
glType [genome|genomephased|exome|rna]
Used for determining if the sample is DNA/RNA/etc. and adding the corresponding tasks to the final workflow. Each sample discovered will take this attribute from the first file encountered for that sample in the config file. -
limsLibraryRecordId
Generated by our LIMS, this allows for the input of data back into the LIMS via a REST-API. -
numberOfReads
Used for validating the number of chunks created during alignment. -
read1Length / read2Length
Used to select the correct STAR indexes. -
readOrientation [inward|outward]
Used to set the strand orientation of RNA assays. Used in conjunction with rnaStrandDirection and rnaStrandType. -
rg values
These are standards set in the SAM/BAM Format Specification:
rgcn - Name of sequencing center producing the read
rgid - Read group identifier.
rgbc - Barcode sequence identifying the sample or library.
rglb - Unique identifier for the library.
rgpl - Platform/technology used to produce the reads.
rgpm - Platform model. Used to configure platform duplicate marking thresholds. Free-form text providing further details of the platform/technology used.
rgpu - Platform unit (e.g., flowcell-barcode.lane for Illumina or slide for SOLiD). Unique identifier.
rgsm - Sample. Use pool name where a pool is being sequenced. -
fraction
Relevant to the TGen naming scheme. See TGen Naming Convention. -
rnaStrandDirection [notapplicable|forward|reverse]
Used during STAR alignment of RNA. -
rnaStrandType [unstranded|stranded]
Assigns the strand orientation of an RNA library -
sampleMergeKey
This is the expected BAM filename and is used to merge data from multiple sequencing lanes or flowcells for data from the same specimen (rgsm) tested with the same assay -
sampleName
This is the expected base FASTQ filename. -
subGroup
Sets where the data file is for tumour or constitutional, changes the analysis of the data file as well as sets the distinction of files during somatic analysis.
Many of the naming structures used are defined by the standardize naming structure used at TGen that ensures all files have a unique but descriptive name. It is designed to support serial collection and multiple collections from difference sources on a single day. Furthermore, sample processing methods can be encoded.
STUDY_PATIENT_VISIT_SOURCE_FRACTION_SubgroupIncrement_ASSAY_LIBRARY
Patient_ID = STUDY_PATIENT
Visit_ID = STUDY_PATIENT_VISIT
Specimen_ID = STUDY_PATIENT_VISIT_SOURCE
Sample_ID = STUDY_PATIENT_VISIT_SOURCE_FRACTION
RG.SM = STUDY_PATIENT_VISIT_SOURCE_FRACTION_SubgroupIncrement (VCF file genotype column header)
sampleMergeKey = STUDY_PATIENT_VISIT_SOURCE_FRACTION_SubgroupIncrement_ASSAY (BAM filename, ensures different assays are not merged together)