Explore Workflows

A workflow for the Broad Institute's best practices gatk4 germline variant calling pipeline. ## __Outputs__ #### Primary Output files: - bqsr2_indels.vcf, filtered and recalibrated indels (IGV browser) - bqsr2_snps.vcf, filtered and recalibrated snps (IGV browser) - bqsr2_snps.ann.vcf, filtered and recalibrated snps with effect annotations #### Secondary Output files: - sorted_dedup_reads.bam, sorted deduplicated alignments (IGV browser) - raw_indels.vcf, first pass indel calls - raw_snps.vcf, first pass snp calls #### Reports: - overview.md (input list, alignment metrics, variant counts) - insert_size_histogram.pdf - recalibration_plots.pdf - snpEff_summary.html ## __Inputs__ #### General Info - Sample short name/Alias: unique name for sample - Experimental condition: condition, variable, etc name (e.g. \"control\" or \"20C 60min\") - Cells: name of cells used for the sample - Catalog No.: vender catalog number if available - BWA index: BWA index sample that contains reference genome FASTA with associated indices. - SNPEFF database: Name of SNPEFF database to use for SNP effect annotation. - Read 1 file: First FASTQ file (generally contains \"R1\" in the filename) - Read 2 file: Paired FASTQ file (generally contains \"R2\" in the filename) #### Advanced - Ploidy: number of copies per chromosome (default should be 2) - SNP filters: see Step 6 Notes: https://gencore.bio.nyu.edu/variant-calling-pipeline-gatk4/ - Indel filters: see Step 7 Notes: https://gencore.bio.nyu.edu/variant-calling-pipeline-gatk4/ #### SNPEFF notes: Get snpeff databases using `docker run --rm -ti gatk4-dev /bin/bash` then running `java -jar $SNPEFF_JAR databases`. Then, use the first column as SNPEFF input (e.g. \"hg38\"). - hg38, Homo_sapiens (USCS), http://downloads.sourceforge.net/project/snpeff/databases/v4_3/snpEff_v4_3_hg38.zip - mm10, Mus_musculus, http://downloads.sourceforge.net/project/snpeff/databases/v4_3/snpEff_v4_3_mm10.zip - dm6.03, Drosophila_melanogaster, http://downloads.sourceforge.net/project/snpeff/databases/v4_3/snpEff_v4_3_dm6.03.zip - Rnor_6.0.86, Rattus_norvegicus, http://downloads.sourceforge.net/project/snpeff/databases/v4_3/snpEff_v4_3_Rnor_6.0.86.zip - R64-1-1.86, Saccharomyces_cerevisiae, http://downloads.sourceforge.net/project/snpeff/databases/v4_3/snpEff_v4_3_R64-1-1.86.zip ### __Data Analysis Steps__ 1. Trimming the adapters with TrimGalore. - This step is particularly important when the reads are long and the fragments are short - resulting in sequencing adapters at the ends of reads. If adapter is not removed the read will not map. TrimGalore can recognize standard adapters, such as Illumina or Nextera/Tn5 adapters. 2. Generate quality control statistics of trimmed, unmapped sequence data 3. Run germline variant calling pipeline, custom wrapper script implementing Steps 1 - 17 of the Broad Institute's best practices gatk4 germline variant calling pipeline (https://gencore.bio.nyu.edu/variant-calling-pipeline-gatk4/) ### __References__ 1. https://gencore.bio.nyu.edu/variant-calling-pipeline-gatk4/ 2. https://gatk.broadinstitute.org/hc/en-us/articles/360035535932-Germline-short-variant-discovery-SNPs-Indels- 3. https://software.broadinstitute.org/software/igv/VCF

https://github.com/datirium/workflows.git

Path: workflows/vc-germline-pe.cwl

Branch/Commit ID: fa4f172486288a1a9d23864f1d6962d85a453e16

https://github.com/common-workflow-language/cwl-utils.git

Path: testdata/workflow_input_format_expr_v1_1.cwl

Branch/Commit ID: 8058c7477097f90205dd7d8481781eb3737ea9c9

https://github.com/common-workflow-language/cwl-v1.2.git

Path: tests/count-lines6-wf.cwl

Branch/Commit ID: 7d7986a6e852ca6e3239c96d3a05dd536c76c903

Note: should be updated The original [BioWardrobe's](https://biowardrobe.com) [PubMed ID:26248465](https://www.ncbi.nlm.nih.gov/pubmed/26248465) **RNA-Seq** basic analysis for a **single-end** experiment. A corresponded input [FASTQ](http://maq.sourceforge.net/fastq.shtml) file has to be provided. Current workflow should be used only with the single-end RNA-Seq data. It performs the following steps: 1. Trim adapters from input FASTQ file 2. Use STAR to align reads from input FASTQ file according to the predefined reference indices; generate unsorted BAM file and alignment statistics file 3. Use fastx_quality_stats to analyze input FASTQ file and generate quality statistics file 4. Use samtools sort to generate coordinate sorted BAM(+BAI) file pair from the unsorted BAM file obtained on the step 1 (after running STAR) 5. Generate BigWig file on the base of sorted BAM file 6. Map input FASTQ file to predefined rRNA reference indices using Bowtie to define the level of rRNA contamination; export resulted statistics to file 7. Calculate isoform expression level for the sorted BAM file and GTF/TAB annotation file using GEEP reads-counting utility; export results to file

https://github.com/datirium/workflows.git

Path: workflows/trim-rnaseq-se-dutp.cwl

Branch/Commit ID: fa4f172486288a1a9d23864f1d6962d85a453e16

SubWorkflow for prodigal. Protein-coding gene prediction for prokaryotic genomes.

https://github.com/EBI-Metagenomics/emg-viral-pipeline.git

Path: cwl/src/Tools/Prodigal/prodigal_swf.cwl

Branch/Commit ID: b0ed3f07c8faced85609287759596ad83e154977

The original [BioWardrobe's](https://biowardrobe.com) [PubMed ID:26248465](https://www.ncbi.nlm.nih.gov/pubmed/26248465) **RNA-Seq** basic analysis for a **paired-end** experiment. A corresponded input [FASTQ](http://maq.sourceforge.net/fastq.shtml) file has to be provided. Current workflow should be used only with the paired-end RNA-Seq data. It performs the following steps: 1. Use STAR to align reads from input FASTQ files according to the predefined reference indices; generate unsorted BAM file and alignment statistics file 2. Use fastx_quality_stats to analyze input FASTQ files and generate quality statistics files 3. Use samtools sort to generate coordinate sorted BAM(+BAI) file pair from the unsorted BAM file obtained on the step 1 (after running STAR) 4. Generate BigWig file on the base of sorted BAM file 5. Map input FASTQ files to predefined rRNA reference indices using Bowtie to define the level of rRNA contamination; export resulted statistics to file 6. Calculate isoform expression level for the sorted BAM file and GTF/TAB annotation file using GEEP reads-counting utility; export results to file

https://github.com/datirium/workflows.git

Path: workflows/rnaseq-pe-dutp.cwl

Branch/Commit ID: fa4f172486288a1a9d23864f1d6962d85a453e16

https://github.com/common-workflow-language/cwl-utils.git

Path: testdata/step_valuefrom5_wf_v1_0.cwl

Branch/Commit ID: 8058c7477097f90205dd7d8481781eb3737ea9c9

https://github.com/common-workflow-language/cwl-utils.git

Path: testdata/workflow_input_sf_expr_array.cwl

Branch/Commit ID: 8058c7477097f90205dd7d8481781eb3737ea9c9

SoupX Estimate ==============

https://github.com/datirium/workflows.git

Path: workflows/soupx.cwl

Branch/Commit ID: fa4f172486288a1a9d23864f1d6962d85a453e16

Super-enhancers, consist of clusters of enhancers that are densely occupied by the master regulators and Mediator. Super-enhancers differ from typical enhancers in size, transcription factor density and content, ability to activate transcription, and sensitivity to perturbation. Use to create stitched enhancers, and to separate super-enhancers from typical enhancers using sequencing data (.bam) given a file of previously identified constituent enhancers (.gff)

https://github.com/datirium/workflows.git

Path: workflows/super-enhancer.cwl

Branch/Commit ID: fa4f172486288a1a9d23864f1d6962d85a453e16