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scdrake offers two pipelines - one for single-sample, and second one for integration of multiple samples (which were processed by the single-sample pipeline before). As for now, each pipeline consists of two subpipelines (referred to as stages), and two stages common to both single-sample and integration pipelines.

A high-level diagram of {scdrake} pipelines.

A more detailed diagram with target structure can be found here.

Each stage has its own config, plus there is a main config for each pipeline. You can read more about configs in a separate vignette("scdrake_config"). Each stage also outputs a report in HTML format with rich graphics.

Advanced users might be interested in looking into source code of scdrake’s plans (files named plans_*.R).

Pipeline steps are mostly based on recommendations given in a great book Orchestrating Single-Cell Analysis with Bioconductor.

Example pipeline output

You can inspect output from the pipeline here.

The used datasets are:

  • PBMC 1k (v3 chemistry, Cell Ranger 3.0.0)
  • PBMC 3k (v2 chemistry, Cell Ranger 1.1.0)

All credits for these datasets go to 10x Genomics. Visit https://www.10xgenomics.com/resources/datasets for more information.


Pipelines

Single-sample pipeline

This is a pipeline for processing a single-sample.

Stages


Integration pipeline

This is a pipeline to integrate multiple samples processed by the single-sample pipeline. Just for clarification, an individual sample is also denoted as a batch.

More information can be found in OSCA

Stages

Stage 02_int_clustering

This stage basically reproduces the clustering and cell type annotation steps in the 02_norm_clustering stage of the single-sample pipeline. The only difference is the user selection of a final integration method which will be used downstream. HVGs, reduced dimensions, and selected markers are already computed in the previous stage (01_integration).


Common stages

Some stages are common to both single-sample and integration pipelines.

Stage cluster_markers

A stage for calculation, visualization and reporting of cell cluster markers (“global markers”).

-> vignette("stage_cluster_markers")

Stage contrasts

A stage for calculation, visualization and reporting of differentially expressed markers (“contrasts”). This stage is basically the same as the cluster_markers stage, but all output is related to individual comparisons of levels of cell groupings. Hence “contrasts”, a term known from bulk RNA-seq where sample groups are compared -> they are put in contrast.

-> vignette("stage_contrasts")


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