Explore Workflows

The original [BioWardrobe's](https://biowardrobe.com) [PubMed ID:26248465](https://www.ncbi.nlm.nih.gov/pubmed/26248465) **ChIP-Seq** basic analysis workflow for a **paired-end** experiment. A [FASTQ](http://maq.sourceforge.net/fastq.shtml) input file has to be provided. The pipeline produces a sorted BAM file alongside with index BAI file, quality statistics of the input FASTQ file, coverage by estimated fragments as a BigWig file, peaks calling data in a form of narrowPeak or broadPeak files, islands with the assigned nearest genes and region type, data for average tag density plot. Workflow starts with step *fastx\_quality\_stats* from FASTX-Toolkit to calculate quality statistics for input FASTQ file. At the same time `bowtie` is used to align reads from input FASTQ file to reference genome *bowtie\_aligner*. The output of this step is an unsorted SAM file which is being sorted and indexed by `samtools sort` and `samtools index` *samtools\_sort\_index*. Depending on workflow’s input parameters indexed and sorted BAM file can be processed by `samtools rmdup` *samtools\_rmdup* to get rid of duplicated reads. If removing duplicates is not required the original BAM and BAI files are returned. Otherwise step *samtools\_sort\_index\_after\_rmdup* repeat `samtools sort` and `samtools index` with BAM and BAI files without duplicates. Next `macs2 callpeak` performs peak calling *macs2\_callpeak* and the next step reports *macs2\_island\_count* the number of islands and estimated fragment size. If the latter is less that 80bp (hardcoded in the workflow) `macs2 callpeak` is rerun again with forced fixed fragment size value (*macs2\_callpeak\_forced*). It is also possible to force MACS2 to use pre set fragment size in the first place. Next step (*macs2\_stat*) is used to define which of the islands and estimated fragment size should be used in workflow output: either from *macs2\_island\_count* step or from *macs2\_island\_count\_forced* step. If input trigger of this step is set to True it means that *macs2\_callpeak\_forced* step was run and it returned different from *macs2\_callpeak* step results, so *macs2\_stat* step should return [fragments\_new, fragments\_old, islands\_new], if trigger is False the step returns [fragments\_old, fragments\_old, islands\_old], where sufix \"old\" defines results obtained from *macs2\_island\_count* step and sufix \"new\" - from *macs2\_island\_count\_forced* step. The following two steps (*bamtools\_stats* and *bam\_to\_bigwig*) are used to calculate coverage from BAM file and save it in BigWig format. For that purpose bamtools stats returns the number of mapped reads which is then used as scaling factor by bedtools genomecov when it performs coverage calculation and saves it as a BEDgraph file whichis then sorted and converted to BigWig format by bedGraphToBigWig tool from UCSC utilities. Step *get\_stat* is used to return a text file with statistics in a form of [TOTAL, ALIGNED, SUPRESSED, USED] reads count. Step *island\_intersect* assigns nearest genes and regions to the islands obtained from *macs2\_callpeak\_forced*. Step *average\_tag\_density* is used to calculate data for average tag density plot from the BAM file.

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

Path: workflows/chipseq-pe.cwl

Branch/Commit ID: bf80c9339d81a78aefb8de661bff998ed86e836e

https://github.com/EBI-Metagenomics/pipeline-v5.git

Path: workflows/subworkflows/final_chunking.cwl

Branch/Commit ID: 4b98d8bf882bc96d924b5d2d4e6d9c188fa7b273

https://github.com/genome/analysis-workflows.git

Path: definitions/pipelines/cle_somatic_exome.cwl

Branch/Commit ID: ae75b938e6e8ae777a55686bbacad824b3c6788c

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

Path: tests/count-lines16-wf.cwl

Branch/Commit ID: 31ec48a8d81ef7c1b2c5e9c0a19e7623efe4a1e2

A CWL workflow for discovering known or novel miRNAs from deep sequencing data using the miRDeep2 tool. The ExoCarta exosome database is also used for identifying exosome-related miRNAs, and TargetScan's organism-specific databases are used for identifying miRNA gene targets. ## __Outputs__ #### Primary Output files: - mirs_known.tsv, detected known mature miRNAs, \"Known miRNAs\" tab - mirs_novel.tsv, detected novel mature miRNAs, \"Novel miRNAs\" tab #### Secondary Output files: - mirs_known_exocarta_deepmirs.tsv, list of detected miRNA also in ExoCarta's exosome database, \"Detected Exosome miRNAs\" tab - mirs_known_gene_targets.tsv, pre-computed gene targets of known mature mirs, downloadable - known_mirs_mature.fa, known mature mir sequences, downloadable - known_mirs_precursor.fa, known precursor mir sequences, downloadable - novel_mirs_mature.fa, novel mature mir sequences, downloadable - novel_mirs_precursor.fa, novel precursor mir sequences, downloadable #### Reports: - overview.md (input list, alignment & mir metrics), \"Overview\" tab - mirdeep2_result.html, summary of mirdeep2 results, \"miRDeep2 Results\" tab ## __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 - Bowtie2 index: Bowtie2 index directory of the reference genome. - Reference Genome FASTA: Reference genome FASTA file to be used for alignment. - Genome short name: Name used for setting organism name, genus, species, and tax ID. - Input FASTQ file: FASTQ file from a single-end miRNA sequencing run. #### Advanced - Adapter: Adapter sequence to be trimmed from miRNA sequence reads. (Default: TCGTAT) - Threads: Number of threads to use for steps that support multithreading (Default: 4). ## Hints & Tips: #### For the identification of novel miRNA candidates, the following may be used as a filtering guideline: 1. miRDeep score > 4 (some authors use 1) 2. not present a match with rfam 3. should present a significant RNAfold (\"yes\") 4. a number of mature reads > 10 5. if applicable, novel mir must be expressed in multiple samples #### For filtering mirbase by organism. | genome | organism | division | name | tree | NCBI-taxid | | ---- | --- | --- | ----------- | ----------- | ----------- | | hg19 | hsa | HSA | Homo sapiens | Metazoa;Bilateria;Deuterostoma;Chordata;Vertebrata;Mammalia;Primates;Hominidae | 9606 | | hg38 | hsa | HSA | Homo sapiens | Metazoa;Bilateria;Deuterostoma;Chordata;Vertebrata;Mammalia;Primates;Hominidae | 9606 | | mm10 | mmu | MMU | Mus musculus | Metazoa;Bilateria;Deuterostoma;Chordata;Vertebrata;Mammalia;Rodentia | 10090 | | rn7 | rno | RNO | Rattus norvegicus | Metazoa;Bilateria;Deuterostoma;Chordata;Vertebrata;Mammalia;Rodentia | 10116 | | dm3 | dme | DME | Drosophila melanogaster | Metazoa;Bilateria;Ecdysozoa;Arthropoda;Hexapoda | 7227 | ## __Data Analysis Steps__ 1. The miRDeep2 Mapper module processes Illumina FASTQ output and maps it to the reference genome. 2. The miRDeep2 miRDeep2 module identifies known and novel (mature and precursor) miRNAs. 3. The ExoCarta database of miRNA found in exosomes is then used to find overlap between mirs_known.tsv and exosome associated miRNAs. 4. Finally, TargetScan organism-specific miRNA gene target database is used to find overlap between mirs_known.tsv and gene targets. ## __References__ 1. https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3245920 2. https://github.com/rajewsky-lab/mirdeep2 3. https://biocontainers.pro/tools/mirdeep2 4. https://www.mirbase.org/ 5. http://exocarta.org/index.html 6. https://www.targetscan.org/vert_80/

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

Path: workflows/mirna-mirdeep2-se.cwl

Branch/Commit ID: 549fac35bf6b8b1c25af0f4f6c3f162c40dc130e

https://github.com/EBI-Metagenomics/pipeline-v5.git

Path: workflows/subworkflows/assembly/CGC-subwf.cwl

Branch/Commit ID: 4b98d8bf882bc96d924b5d2d4e6d9c188fa7b273

https://github.com/genome/analysis-workflows.git

Path: definitions/subworkflows/molecular_qc.cwl

Branch/Commit ID: aba52e94b6d7470132d3c092c26d67e29d615300

https://github.com/ncbi/pgap.git

Path: task_types/tt_univec_wnode.cwl

Branch/Commit ID: 2d851682ba1bf2aaaacb3677253b55ceb59c8568

https://github.com/d3b-center/canine-dev.git

Path: subworkflows/canine_manta_module.cwl

Branch/Commit ID: 462aaebbd442e84ea101b45b716df0174b88512e

https://github.com/common-workflow-language/cwltool.git

Path: tests/wf/conflict.cwl

Branch/Commit ID: 4fd5ca5a927594c361a9320d5331b326d06cecd3

Packed ID: main