Explore Workflows

Cellranger Reanalyze ====================

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

Path: workflows/cellranger-reanalyze.cwl

Branch/Commit ID: 3042f91bf93bd91c0f420c53aaf10874967e8ddc

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

Path: tests/sum-wf.cwl

Branch/Commit ID: 1f3ef888d9ef2306c828065c460c1800604f0de4

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

Path: tests/count-lines8-wf-noET.cwl

Branch/Commit ID: 1f3ef888d9ef2306c828065c460c1800604f0de4

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: 93b844a80f4008cc973ea9b5efedaff32a343895

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

Path: tests/scatter-valuefrom-wf1.cwl

Branch/Commit ID: ea9f8634e41824ac3f81c3dde698d5f0eef54f1b

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

Path: definitions/subworkflows/duplex_alignment.cwl

Branch/Commit ID: e0b3c76e38630fb6234414b5adebfb6a4fb23117

Copy fasta_file file to the folder and run run bismark_genome_preparation script to prepare indices for Bismark Methylation Analysis. Bowtie2 aligner is used by default. The name of the output indices folder is equal to the genome input.

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

Path: workflows/bismark-index.cwl

Branch/Commit ID: 7ced5a5259dbd8b3fc64456beaeffd44f4a24081

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

Path: task_types/tt_kmer_build_tree.cwl

Branch/Commit ID: ce433f771ebf5677c9f40858e2ae91b1a7e75d30

Calculates the rescue ratio (see Gabe's protocols paper), given two eCLIP IP samples and 2 size-matched input samples. Also returns the reproducible peaks given these two samples. This is different from the 1input workflow in that each INPUT is first merged together and is used downstream instead of the 1input version, which remains unmodified. Merged inputs are NOT used in calculating true reproducible peaks.

https://github.com/YeoLab/merge_peaks.git

Path: cwl/wf_rescue_ratio_2inputs.cwl

Branch/Commit ID: 55f4f4f9c10a09ce03c5c531dd176e6080118977

https://github.com/ebi-wp/webservice-cwl.git

Path: workflows/workflow-fasta-pratt.cwl

Branch/Commit ID: 7c2c01c23d7a68a4f0c608881280576d65a01325