Explore Workflows

A workflow for processing a single 16S sample via a QIIME2 pipeline. ## __Outputs__ #### Output files: - overview.md, list of inputs - demux.qzv, summary visualizations of imported data - alpha-rarefaction.qzv, plot of OTU rarefaction - taxa-bar-plots.qzv, relative frequency of taxomonies barplot ## __Inputs__ #### General Info - Sample short name/Alias: Used for samplename in downstream analyses. Ensure this is the same name used in the metadata samplesheet. - Environment: where the sample was collected - Catalog No.: catalog number if available (optional) - Read 1 FASTQ file: Read 1 FASTQ file from a paired-end sequencing run. - Read 2 FASTQ file: Read 2 FASTQ file that pairs with the input R1 file. - Trim 5' of R1: Recommended if adapters are still on the input sequences. Trims the first J bases from the 5' end of each forward read. - Trim 5' of R2: Recommended if adapters are still on the input sequences. Trims the first K bases from the 5' end of each reverse read. - Truncate 3' of R1: Recommended if quality drops off along the length of the read. Clips the forward read starting M bases from the 5' end (before trimming). - Truncate 3' of R2: Recommended if quality drops off along the length of the read. Clips the reverse read starting N bases from the 5' end (before trimming). - Threads: Number of threads to use for steps that support multithreading. ### __Data Analysis Steps__ 1. Generate FASTX quality statistics for visualization of unmapped, raw FASTQ reads. 2. Import the data, make a qiime artifact (demux.qza), and summary visualization 3. Denoising will detect and correct (where possible) Illumina amplicon sequence data. This process will additionally filter any phiX reads (commonly present in marker gene Illumina sequence data) that are identified in the sequencing data, and will filter chimeric sequences. 4. Generate a phylogenetic tree for diversity analyses and rarefaction processing and plotting. 5. Taxonomy classification of amplicons. Performed using a Naive Bayes classifier trained on the Greengenes2 database \"gg_2022_10_backbone_full_length.nb.qza\". ### __References__ 1. Bolyen E, Rideout JR, Dillon MR, Bokulich NA, Abnet CC, Al-Ghalith GA, Alexander H, Alm EJ, Arumugam M, Asnicar F, Bai Y, Bisanz JE, Bittinger K, Brejnrod A, Brislawn CJ, Brown CT, Callahan BJ, Caraballo-Rodríguez AM, Chase J, Cope EK, Da Silva R, Diener C, Dorrestein PC, Douglas GM, Durall DM, Duvallet C, Edwardson CF, Ernst M, Estaki M, Fouquier J, Gauglitz JM, Gibbons SM, Gibson DL, Gonzalez A, Gorlick K, Guo J, Hillmann B, Holmes S, Holste H, Huttenhower C, Huttley GA, Janssen S, Jarmusch AK, Jiang L, Kaehler BD, Kang KB, Keefe CR, Keim P, Kelley ST, Knights D, Koester I, Kosciolek T, Kreps J, Langille MGI, Lee J, Ley R, Liu YX, Loftfield E, Lozupone C, Maher M, Marotz C, Martin BD, McDonald D, McIver LJ, Melnik AV, Metcalf JL, Morgan SC, Morton JT, Naimey AT, Navas-Molina JA, Nothias LF, Orchanian SB, Pearson T, Peoples SL, Petras D, Preuss ML, Pruesse E, Rasmussen LB, Rivers A, Robeson MS, Rosenthal P, Segata N, Shaffer M, Shiffer A, Sinha R, Song SJ, Spear JR, Swafford AD, Thompson LR, Torres PJ, Trinh P, Tripathi A, Turnbaugh PJ, Ul-Hasan S, van der Hooft JJJ, Vargas F, Vázquez-Baeza Y, Vogtmann E, von Hippel M, Walters W, Wan Y, Wang M, Warren J, Weber KC, Williamson CHD, Willis AD, Xu ZZ, Zaneveld JR, Zhang Y, Zhu Q, Knight R, and Caporaso JG. 2019. Reproducible, interactive, scalable and extensible microbiome data science using QIIME 2. Nature Biotechnology 37: 852–857. https://doi.org/10.1038/s41587-019-0209-9

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

Path: workflows/qiime2-sample-pe.cwl

Branch/Commit ID: 93b844a80f4008cc973ea9b5efedaff32a343895

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

Path: testdata/count-lines7-wf_v1_0.cwl

Branch/Commit ID: b76b039edb62dea76c43f173848cdc57e4b4aab7

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

Path: testdata/record-output-wf_v1_2.cwl

Branch/Commit ID: b76b039edb62dea76c43f173848cdc57e4b4aab7

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

Path: testdata/record-output-wf_v1_0.cwl

Branch/Commit ID: b76b039edb62dea76c43f173848cdc57e4b4aab7

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

Path: definitions/subworkflows/gatk_haplotypecaller_iterator.cwl

Branch/Commit ID: 39ac49f5d080bbb6bfa97246f46a5b621254f622

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

Path: testdata/md5sum.cwl

Branch/Commit ID: b76b039edb62dea76c43f173848cdc57e4b4aab7

Interval overlapping alignments counts ====================================== Reports the count of alignments from multiple samples that overlap specific intervals.

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

Path: workflows/bedtools-multicov.cwl

Branch/Commit ID: 93b844a80f4008cc973ea9b5efedaff32a343895

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: 93b844a80f4008cc973ea9b5efedaff32a343895

DESeq2 Multi-factor Analysis Runs DeSeq2 multi-factor analysis with manual control over major parameters

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

Path: workflows/deseq-multi-factor.cwl

Branch/Commit ID: 93b844a80f4008cc973ea9b5efedaff32a343895

This workflow indexes the input reference FASTA with bwa, and generates faidx and dict file using samtools. This index sample can then be used as input into the germline variant calling workflow, or others that may include this workflow as an upstream source. ### __Inputs__ - FASTA file of the reference genome that will be indexed. ### __Outputs__ - Directory containing the original FASTA, faidx, dict, and bwa index files. - stdout log file (output in Overview tab as well) - stderr log file ### __Data Analysis Steps__ 1. cwl calls dockercontainer robertplayer/scidap-gatk4 to index reference FASTA with bwa, and generates faidx and dict files using samtools ### __References__ - Li, H., & Durbin, R. (2009). Fast and accurate short read alignment with Burrows–Wheeler transform. Bioinformatics, 25(14), 1754–1760.

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

Path: workflows/bwa-index.cwl

Branch/Commit ID: 93b844a80f4008cc973ea9b5efedaff32a343895