Cluster 3k PBMCs with Seurat - Workflow - SCTransform Version

single-cell-scrna-seurat-pbmc3k/seurat-pbmc-workflow-sct

Author(s)
Marisa Loach
version Version
1
last_modification Last updated
Jan 23, 2025
license License
CC-BY-4.0
galaxy-tags Tags
name:singlecell
name:seurat

Features
Tutorial
hands_on Clustering 3K PBMCs with Seurat
workflow Other workflows associated with this material
Workflow Testing
Tests: ✅
Results: Not yet automated
FAIRness purl PURL
https://gxy.io/GTN:W00306
RO-Crate logo with flask Download Workflow RO-Crate Workflowhub cloud with gears logo View on (Dev) WorkflowHub
Launch in Tutorial Mode question
galaxy-download Download
flowchart TD
  0["ℹ️ Input Dataset\nmatrix.mtx"];
  style 0 stroke:#2c3143,stroke-width:4px;
  1["ℹ️ Input Dataset\ngenes.tsv"];
  style 1 stroke:#2c3143,stroke-width:4px;
  2["ℹ️ Input Dataset\nbarcodes.tsv"];
  style 2 stroke:#2c3143,stroke-width:4px;
  3["ℹ️ Input Dataset\nCanonical Markers"];
  style 3 stroke:#2c3143,stroke-width:4px;
  4["Input 3k PBMCs"];
  2 -->|output| 4;
  1 -->|output| 4;
  0 -->|output| 4;
  13a7e18a-3194-4ba9-adce-7e934896f807["Output\nInput 3k PBMC"];
  4 --> 13a7e18a-3194-4ba9-adce-7e934896f807;
  style 13a7e18a-3194-4ba9-adce-7e934896f807 stroke:#2c3143,stroke-width:4px;
  5["Seurat Data Management"];
  4 -->|rds_out| 5;
  6["Mitochondrial Annotations"];
  4 -->|rds_out| 6;
  7["Seurat Visualize"];
  6 -->|rds_out| 7;
  8["Seurat Visualize"];
  6 -->|rds_out| 8;
  9["Seurat Visualize"];
  6 -->|rds_out| 9;
  10["Filtered Dataset"];
  6 -->|rds_out| 10;
  11["Preprocessed Data"];
  10 -->|rds_out| 11;
  1ec75d26-22af-4d32-bd26-ef5a691c7612["Output\nPreprocessed Data"];
  11 --> 1ec75d26-22af-4d32-bd26-ef5a691c7612;
  style 1ec75d26-22af-4d32-bd26-ef5a691c7612 stroke:#2c3143,stroke-width:4px;
  12["Seurat Data Management"];
  11 -->|rds_out| 12;
  13["Seurat Visualize"];
  11 -->|rds_out| 13;
  14["PCA Results"];
  11 -->|rds_out| 14;
  15["Seurat Visualize"];
  14 -->|rds_out| 15;
  16["Seurat Visualize"];
  14 -->|rds_out| 16;
  17["Seurat Visualize"];
  14 -->|rds_out| 17;
  18["Seurat Visualize"];
  14 -->|rds_out| 18;
  19["Seurat Visualize"];
  14 -->|rds_out| 19;
  20["Seurat Visualize"];
  14 -->|rds_out| 20;
  21["Seurat Visualize"];
  14 -->|rds_out| 21;
  22["Seurat Find Clusters"];
  14 -->|rds_out| 22;
  23["Seurat Find Clusters"];
  22 -->|rds_out| 23;
  24["UMAP Results"];
  23 -->|rds_out| 24;
  25["Seurat Visualize"];
  24 -->|rds_out| 25;
  26["Seurat Visualize"];
  24 -->|rds_out| 26;
  27["Seurat Visualize"];
  24 -->|rds_out| 27;
  28["DE Markers"];
  24 -->|rds_out| 28;
  29["Seurat Visualize"];
  24 -->|rds_out| 29;
  30["Seurat Visualize"];
  24 -->|rds_out| 30;
  31["Seurat Visualize"];
  24 -->|rds_out| 31;
  32["Seurat Visualize"];
  24 -->|rds_out| 32;
  33["Annotated Clusters"];
  24 -->|rds_out| 33;
  8d42442d-ba15-41f6-bbd7-44b551c1bccf["Output\nAnnotated Clusters"];
  33 --> 8d42442d-ba15-41f6-bbd7-44b551c1bccf;
  style 8d42442d-ba15-41f6-bbd7-44b551c1bccf stroke:#2c3143,stroke-width:4px;
  34["DE Markers Tabular"];
  28 -->|markers_tabular| 34;
  35["Seurat Visualize"];
  33 -->|rds_out| 35;
  36["Seurat Visualize"];
  3 -->|output| 36;
  33 -->|rds_out| 36;
  37["Input DE Markers"];
  34 -->|tabular| 37;
  d2e05a88-9176-4650-9ad7-285a63ae86f9["Output\nInput DE Markers"];
  37 --> d2e05a88-9176-4650-9ad7-285a63ae86f9;
  style d2e05a88-9176-4650-9ad7-285a63ae86f9 stroke:#2c3143,stroke-width:4px;
  38["Seurat Visualize"];
  37 -->|out_file1| 38;
  33 -->|rds_out| 38;

Inputs

Input Label
matrix.mtx matrix.mtx
genes.tsv genes.tsv
barcodes.tsv barcodes.tsv
Canonical Markers Canonical Markers

Outputs

From Output Label
toolshed.g2.bx.psu.edu/repos/iuc/seurat_create/seurat_create/5.0+galaxy1 Seurat Create Input 3k PBMCs
toolshed.g2.bx.psu.edu/repos/iuc/seurat_preprocessing/seurat_preprocessing/5.0+galaxy0 Seurat Preprocessing Preprocessed Data
toolshed.g2.bx.psu.edu/repos/iuc/seurat_data/seurat_data/5.0+galaxy0 Seurat Data Management Annotated Clusters
Cut1 Cut Input DE Markers

Tools

Tool Links
Cut1
csv_to_tabular
toolshed.g2.bx.psu.edu/repos/iuc/seurat_clustering/seurat_clustering/5.0+galaxy0 View in ToolShed
toolshed.g2.bx.psu.edu/repos/iuc/seurat_create/seurat_create/5.0+galaxy1 View in ToolShed
toolshed.g2.bx.psu.edu/repos/iuc/seurat_data/seurat_data/5.0+galaxy0 View in ToolShed
toolshed.g2.bx.psu.edu/repos/iuc/seurat_plot/seurat_plot/5.0+galaxy0 View in ToolShed
toolshed.g2.bx.psu.edu/repos/iuc/seurat_preprocessing/seurat_preprocessing/5.0+galaxy0 View in ToolShed
toolshed.g2.bx.psu.edu/repos/iuc/seurat_reduce_dimension/seurat_reduce_dimension/5.0+galaxy0 View in ToolShed

To use these workflows in Galaxy you can either click the links to download the workflows, or you can right-click and copy the link to the workflow which can be used in the Galaxy form to import workflows.

Importing into Galaxy

Below are the instructions for importing these workflows directly into your Galaxy server of choice to start using them!
Hands-on: Importing a workflow
  • Click on Workflow on the top menu bar of Galaxy. You will see a list of all your workflows.
  • Click on galaxy-upload Import at the top-right of the screen
  • Provide your workflow
    • Option 1: Paste the URL of the workflow into the box labelled “Archived Workflow URL”
    • Option 2: Upload the workflow file in the box labelled “Archived Workflow File”
  • Click the Import workflow button

Below is a short video demonstrating how to import a workflow from GitHub using this procedure:

Video: Importing a workflow from URL

Version History

Version Commit Time Comments
6 7337cb11a 2025-01-08 10:16:27 Correcting file name 4
5 ed2e0fc88 2025-01-08 10:14:23 Deleting old file
4 6a4c55c1f 2024-12-04 14:03:00 Changing a few parameters
3 4ddf97dbc 2024-10-31 14:04:51 Fixing errors in Seurat_PBMC_Workflow_SCT.ga
2 6bcca5b8d 2024-10-31 13:27:31 Adding workflows with workflow tests
1 8f1616f52 2024-10-29 10:15:02 Shorter name for workflow 2

For Admins

Installing the workflow tools

wget https://training.galaxyproject.org/training-material/topics/single-cell/tutorials/scrna-seurat-pbmc3k/workflows/Seurat_PBMC_Workflow_SCT.ga -O workflow.ga
workflow-to-tools -w workflow.ga -o tools.yaml
shed-tools install -g GALAXY -a API_KEY -t tools.yaml
workflow-install -g GALAXY -a API_KEY -w workflow.ga --publish-workflows