Workflow: scRNA-seq pipeline using Salmon and Alevin

Fetched 2023-09-16 08:36:00 GMT
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Inputs

ID Type Title Doc
assay String scRNA-seq assay
threads Integer Number of threads for Salmon
fastq_dir Directory[] Directory containing FASTQ files
keep_all_barcodes Boolean (Optional)
expected_cell_count Integer (Optional)
visium_probe_set_version Integer (Optional)

Steps

ID Runs Label Doc
fastqc
steps/fastqc.cwl (CommandLineTool)
Runs fastQC on each fastq file in fastq directory
quantification
steps/quantification.cwl (CommandLineTool)
Run Salmon Alevin tool on FASTQ input
adjust_barcodes
steps/adjust-barcodes.cwl (CommandLineTool)
Assay-specific adjustment of cell barcodes
scanpy_analysis
steps/scanpy-analysis.cwl (CommandLineTool)
Dimensionality reduction and clustering
squidpy_analysis
steps/squidpy-analysis.cwl (CommandLineTool)
Dimensionality reduction and clustering
compute_qc_results
steps/compute-qc-metrics.cwl (CommandLineTool)
Compute QC metrics

Outputs

ID Type Label Doc
qc_report File Quality control report in JSON format
umap_plot File UMAP dimensionality reduction plot
fastqc_dir Directory[] Directory of FastQC output files, mirroring input directory structure
ripley_plot File
spatial_plot File (Optional) Slide-seq bead plot, colored by Leiden cluster
dispersion_plot File Gene expression dispersion plot
count_matrix_h5ad File Unfiltered count matrix from BWA and umi_tools, converted to H5AD, spliced and unspliced counts
scanpy_qc_results File Quality control metrics from Scanpy
umap_density_plot File UMAP dimensionality reduction plot, colored by cell density
co_occurrence_plot File
filtered_data_h5ad File Full data set of filtered results

Full data set of filtered results: expression matrix, coordinates in dimensionality-reduced space (PCA and UMAP), cluster assignments via the Leiden algorithm, and marker genes for one cluster vs. rest

squidpy_spatial_plot File (Optional)
centrality_scores_plot File
squidpy_annotated_h5ad File
interaction_matrix_plot File
marker_gene_plot_logreg File Cluster marker genes, logreg method
marker_gene_plot_t_test File Cluster marker genes, t-test
neighborhood_enrichment_plot File
Permalink: https://w3id.org/cwl/view/git/f03aa8543a4d13aac6419953f5d17f9eda6c8c17/pipeline.cwl