Explore Workflows

Cell Ranger Count (RNA) Quantifies single-cell gene expression of the sequencing data from a single 10x Genomics library. The results of this workflow are primarily used in either “Single-Cell RNA-Seq Filtering Analysis” or “Cell Ranger Aggregate (RNA, RNA+VDJ)” pipelines.

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

Path: workflows/single-cell-preprocess-cellranger.cwl

Branch/Commit ID: cc6fa135d04737fdde3b4414d6e214cf8c812f6e

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

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

Path: workflows/soupx.cwl

Branch/Commit ID: 00ea05e22788029370898fd4c17798b11edf0e57

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: cc6fa135d04737fdde3b4414d6e214cf8c812f6e

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

Path: tests/wf/paramref_arguments_self.cwl

Branch/Commit ID: e6a1e1f3a3b3168028bd19aaf465826fa276a35b

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

Path: spurious_annot/wf_spurious_annot_pass2.cwl

Branch/Commit ID: 4b73bfeb967ee9f57a0410276f7c39e784f0846f

Runs RNA-Seq BioWardrobe basic analysis with strand specific pair-end data file.

https://github.com/Barski-lab/workflows.git

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

Branch/Commit ID: 80d64741638b14de5cf58236b6d6d99713c62086

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

Path: task_types/tt_format_rrnas_from_seq_entry.cwl

Branch/Commit ID: 23f0ee7a36649ab37cabdd9277b7c82d098be79c

Runs ChIP-Seq BioWardrobe basic analysis with paired-end input data files.

https://github.com/Barski-lab/workflows.git

Path: workflows/chipseq-pe.cwl

Branch/Commit ID: 80d64741638b14de5cf58236b6d6d99713c62086

A basic differential methylation analysis workflow using BismarkCov formatted bed files as input to the RnBeads tool. Analysis is conducted on region and sites levels according to the sample groups specified by user (limited to 2 conditions in this workflow implementation). See report html files for detailed descriptions of analyses and results interpretation. ### __Inputs__ *General Info:* - Experiment short name/Alias* - a unique name for the sample (e.g. what was used on tubes while processing it) - Condition 1 name - name defining condition/group 1 - Condition 2 name - name defining condition/group 2 - Bismark coverage files* for condition1 - minumum of 2 is required for analysis - Bismark coverage files* for condition2 - minumum of 2 is required for analysis - Sample genome - available options: hg19, hg38, mm9, mm10, rn5 - Genome type - indicate mismark index used for upstream samples (input for conditions 1 and 2) *Advanced:* - Number of threads for steps that support multithreading - default set to `4` *[BismarkCov formatted bed](https://www.bioinformatics.babraham.ac.uk/projects/bismark/Bismark_User_Guide.pdf): The genome-wide cytosine report (optional) is tab-delimited in the following format (1-based coords): <chromosome> <position> <strand> <count methylated> <count unmethylated> <C-context> <trinucleotide context> ### __Outputs__ Intermediate and final downloadable outputs include: - sig_dm_sites.bed ([bed for IGV](https://genome.ucsc.edu/FAQ/FAQformat.html#format1); sig diff meth sites) - sig_dm_sites_annotated.tsv (tsv for TABLE; for each site above, closest single gene annotation) - Site_id, unique indentifer per methylated site - Site_Chr, chromosome of methylated site - Site_position, 1-based position in chr of methylated site - Site_strand, strand of methylated site - Log2_Meth_Quotient, log2 of the quotient in methylation: log2((mean.g1+epsilon)/(mean.g2+epsilon)), where epsilon:=0.01. In case of paired analysis, it is the mean of the pairwise quotients. - FDR, adjusted p-values, all <0.10 assumed to be significant - Coverage_score, value between 0-1000 reflects strength of mean coverage difference between conditions and equals [1000-(1000/(meancov_g1-meancov_g2)^2](https://www.wolframalpha.com/input?i=solve+1000-%281000%2F%28x%5E2%29%29), if meancov_g1-meancov_g2==0, score=0, elif score<1==1, else score - meancov_g1, mean coverage of condition1 - meancov_g2, mean coverage of condition2 - refSeq_id, RefSeq gene id - Gene_id, gene symbol - Chr, gene chromosome - txStart, gene transcription start position - tsEnd, gene transcription end position - txStrand, gene strand - stdout and stderr log files - Packaged RnBeads reports directory (reports.tar.gz) contains: reports/ ├── configuration ├── data_import.html ├── data_import_data ├── data_import_images ├── data_import_pdfs ├── differential_methylation.html ├── differential_methylation_data ├── differential_methylation_images ├── differential_methylation_pdfs ├── preprocessing.html ├── preprocessing_data ├── preprocessing_images ├── preprocessing_pdfs ├── quality_control.html ├── quality_control_data ├── quality_control_images ├── quality_control_pdfs ├── tracks_and_tables.html ├── tracks_and_tables_data ├── tracks_and_tables_images └── tracks_and_tables_pdfs Reported methylation is in the form of regions (genes, promoters, cpg, tiling) and specific sites: - genes - Ensembl gene definitions are downloaded using the biomaRt package. - promoters - A promoter is defined as the region spanning 1,500 bases upstream and 500 bases downstream of the transcription start site of the corresponding gene - cpg - the CpG islands from the UCSC Genome Browser - tiling - a window size of 5 kilobases are defined over the whole genome - sites - all cytosines in the context of CpGs in the respective genome ### __Data Analysis Steps__ 1. generate sample sheet with associated conditions for testing in RnBeads 2. setup rnbeads analyses in R, and run differential methylation analysis 3. process output diffmeth files for regions and sites 4. find single closest gene annotations for all significantly diffmeth sites 5. package and save rnbeads report directory 6. clean up report dir for html outputs ### __References__ - https://rnbeads.org/materials/example_3/differential_methylation.html - Makambi, K. (2003) Weighted inverse chi-square method for correlated significance tests. Journal of Applied Statistics, 30(2), 225234 - https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4216143/ - Assenov Y, Müller F, Lutsik P, Walter J, Lengauer T, Bock C. Comprehensive analysis of DNA methylation data with RnBeads. Nat Methods. 2014 Nov;11(11):1138-1140. doi: 10.1038/nmeth.3115. Epub 2014 Sep 28. PMID: 25262207; PMCID: PMC4216143.

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

Path: workflows/diffmeth.cwl

Branch/Commit ID: cc6fa135d04737fdde3b4414d6e214cf8c812f6e

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

Path: task_types/tt_gcaccess_from_list.cwl

Branch/Commit ID: ac387721a55fd91df3dcdf16e199354618b136d1