Mercurial > repos > iuc > scater_create_qcmetric_ready_sce
diff README.md @ 2:b834074a9aff draft
"planemo upload for repository https://github.com/galaxyproject/tools-iuc/tree/master/tools/scater commit 154318f74839a4481c7c68993c4fb745842c4cce"
author | iuc |
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date | Thu, 09 Sep 2021 12:23:11 +0000 |
parents | fd808de478b1 |
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--- a/README.md Tue Sep 03 14:26:31 2019 -0400 +++ b/README.md Thu Sep 09 12:23:11 2021 +0000 @@ -1,20 +1,20 @@ # Wrappers for Scater -This code wraps a number of [scater](https://bioconductor.org/packages/release/bioc/html/scater.html) functions as Galaxy wrappers. Briefly, the `scater-create-qcmetric-ready-sce` tool takes a sample gene expression matrix (usually read-counts) and a cell annotation file, creates a [SingleCellExperiment](https://bioconductor.org/packages/release/bioc/html/SingleCellExperiment.html) object and runs scater's `calculateQCMetrics` function (using other supplied files such as ERCC's and mitochondrial gene features). +This code wraps a number of [scater](https://bioconductor.org/packages/release/bioc/html/scater.html) and [scuttle](https://bioconductor.org/packages/3.13/bioc/html/scuttle.html) functions as Galaxy wrappers. Briefly, the `scater-create-qcmetric-ready-sce` tool takes a sample gene expression matrix (usually read-counts) and a cell annotation file, creates a [SingleCellExperiment](https://bioconductor.org/packages/release/bioc/html/SingleCellExperiment.html) object and runs scater's `calculateQCMetrics` function (using other supplied files such as ERCC's and mitochondrial gene features). Various filter scripts are provided, along with some plotting functions for QC. ## Typical workflow 1. Read in data with `scater-create-qcmetric-ready-sce`. -2. Visualise it.\ +2. Visualise it. Take a look at the distribution of library sizes, expressed features and mitochondrial genes with `scater-plot-dist-scatter`. - Then look at the distibution of genes across cells with `scater-plot-exprs-freq`. + 3. Guided by the plots, filter the data with `scater-filter`.\ You can either manually filter with user-defined parameters or use PCA to automatically removes outliers. 4. Visualise data again to see how the filtering performed using `scater-plot-dist-scatter`.\ Decide if you're happy with the data. If not, try increasing or decreasing the filtering parameters. -5. Normalise data with `scater-normalize`. + 6. Investigate other confounding factors.\ Plot the data (using PCA) and display various annotated properties of the cells using `scater-plot-pca`. @@ -51,10 +51,6 @@ --- -`scater-plot-exprs-freq.R` -Plots mean expression vs % of expressing cells and provides information as to the number of genes expressed in 50% and 25% of cells. - ---- `scater-pca-filter.R` Takes SingleCellExperiment object (from Loom file) and automatically removes outliers from data using PCA. Save the filtered SingleCellExperiment object in Loom format. @@ -74,18 +70,18 @@ --- -`scater-normalize.R` -Compute log-normalized expression values from count data in a SingleCellExperiment object, using the size factors stored in the object. Save the normalised SingleCellExperiment object in Loom format. +`scater-plot-pca.R` +PCA plot of a SingleCellExperiment object. The options `-c`, `-p`, and `-s` all refer to cell annotation features. These are the column headers of the `-c` option used in `scater-create-qcmetric-ready-sce.R`. ``` -./scater-normalize.R -i test-data/scater_manual_filtered.loom -o test-data/scater_man_filtered_normalised.loom +./scater-plot-pca.R -i test-data/scater_qcready.loom -c Treatment -p Mutation_Status -o test-data/scater_pca_plot.pdf ``` --- -`scater-plot-pca.R` -PCA plot of a normalised SingleCellExperiment object (produced with `scater-normalize.R`). The options `-c`, `-p`, and `-s` all refer to cell annotation features. These are the column headers of the `-c` option used in `scater-create-qcmetric-ready-sce.R`. +`scater-plot-tsne.R` +t-SNE plot of a SingleCellExperiment object. The options `-c`, `-p`, and `-s` all refer to cell annotation features. These are the column headers of the `-c` option used in `scater-create-qcmetric-ready-sce.R`. ``` -./scater-plot-pca.R -i test-data/scater_man_filtered_normalised.loom -c Treatment -p Mutation_Status -o test-data/scater_pca_plot.pdf +./scater-plot-tsne.R -i test-data/scater_qcready.loom -c Treatment -p Mutation_Status -o test-data/scater_tsne_plot.pdf ```