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: 1f68f6aa0d71d8f90a7ae37a1c7db4580e199b68

Single-Cell ATAC-Seq Cluster Analysis Clusters cells by similarity of chromatin accessibility data from the outputs of the “Single-Cell ATAC-Seq Dimensionality Reduction Analysis” pipeline. The results of this workflow are used in the “Single-Cell Manual Cell Type Assignment”, “Single-Cell ATAC-Seq Differential Accessibility Analysis”, and “Single-Cell ATAC-Seq Genome Coverage” pipelines.

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

Path: workflows/sc-atac-cluster.cwl

Branch/Commit ID: 1f68f6aa0d71d8f90a7ae37a1c7db4580e199b68

rna / protein - qc, preprocess, filter, annotation, index, abundance

https://github.com/MG-RAST/pipeline.git

Path: CWL/Workflows/wgs-noscreen-fasta.workflow.cwl

Branch/Commit ID: 2addcde0f4c1c8547f7f3906c2523cded23e9869

GSEAPY: Gene Set Enrichment Analysis in Python ============================================== Gene Set Enrichment Analysis is a computational method that determines whether an a priori defined set of genes shows statistically significant, concordant differences between two biological states (e.g. phenotypes). GSEA requires as input an expression dataset, which contains expression profiles for multiple samples. While the software supports multiple input file formats for these datasets, the tab-delimited GCT format is the most common. The first column of the GCT file contains feature identifiers (gene ids or symbols in the case of data derived from RNA-Seq experiments). The second column contains a description of the feature; this column is ignored by GSEA and may be filled with “NA”s. Subsequent columns contain the expression values for each feature, with one sample's expression value per column. It is important to note that there are no hard and fast rules regarding how a GCT file's expression values are derived. The important point is that they are comparable to one another across features within a sample and comparable to one another across samples. Tools such as DESeq2 can be made to produce properly normalized data (normalized counts) which are compatible with GSEA. Documents ============================================== - GSEA Home Page: https://www.gsea-msigdb.org/gsea/index.jsp - Results Interpretation: https://www.gsea-msigdb.org/gsea/doc/GSEAUserGuideTEXT.htm#_Interpreting_GSEA_Results - GSEA User Guide: https://gseapy.readthedocs.io/en/latest/faq.html - GSEAPY Docs: https://gseapy.readthedocs.io/en/latest/introduction.html References ============================================== - Subramanian, Tamayo, et al. (2005, PNAS), https://www.pnas.org/content/102/43/15545 - Mootha, Lindgren, et al. (2003, Nature Genetics), http://www.nature.com/ng/journal/v34/n3/abs/ng1180.html - Chen EY, Tan CM, Kou Y, Duan Q, Wang Z, Meirelles GV, Clark NR, Ma'ayan A. Enrichr: interactive and collaborative HTML5 gene list enrichment analysis tool. BMC Bioinformatics. 2013; 128(14). - Kuleshov MV, Jones MR, Rouillard AD, Fernandez NF, Duan Q, Wang Z, Koplev S, Jenkins SL, Jagodnik KM, Lachmann A, McDermott MG, Monteiro CD, Gundersen GW, Ma'ayan A. Enrichr: a comprehensive gene set enrichment analysis web server 2016 update. Nucleic Acids Research. 2016; gkw377 . - Xie Z, Bailey A, Kuleshov MV, Clarke DJB., Evangelista JE, Jenkins SL, Lachmann A, Wojciechowicz ML, Kropiwnicki E, Jagodnik KM, Jeon M, & Ma’ayan A. Gene set knowledge discovery with Enrichr. Current Protocols, 1, e90. 2021. doi: 10.1002/cpz1.90

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

Path: workflows/gseapy.cwl

Branch/Commit ID: 1f68f6aa0d71d8f90a7ae37a1c7db4580e199b68

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

Path: tests/scatter-valuefrom-wf5.cwl

Branch/Commit ID: c7c97715b400ff2194aa29fc211d3401cea3a9bf

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

Path: task_types/tt_kmer_build_tree.cwl

Branch/Commit ID: 008a090fb1938fbb393494ac8fcb219f0d9f5295

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

Path: task_types/tt_ani_top_n.cwl

Branch/Commit ID: 8a8fffb78b1e327ba0da51840ac8acc0c218d611

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

Path: task_types/tt_blastp_wnode_naming.cwl

Branch/Commit ID: 68b828ac482956a03325623d817780986f34fb31

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

Path: tests/scatter-valuefrom-wf6.cwl

Branch/Commit ID: 1f3ef888d9ef2306c828065c460c1800604f0de4

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

Path: cwltool/schemas/v1.0/v1.0/scatter-valuefrom-wf5.cwl

Branch/Commit ID: 2256a30d0c1365b30e0a7338fb883c74674fcd25