Initial processing using RaceID

performs filtering, normalisation, and confounder removal to generate a normalised and filtered count matrix of single-cell RNA data (Galaxy Version 3.1)

Tool Parameters

Please provide a value for this option.
* required
Filtering
*
The minimum total transcripts required. Cells with less than mintotal transcripts are filtered out.
*
The minimum required transcript counts of a gene in the minimum number of cells (below)
*
The minumum number of cells for gene expression to be counted
By default features are counted if they are above zero, but RaceID adds 0.1 to all counts after normalisation to create a non-zero dataset.
Use Defaults?

Additional Options

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RaceID3

RaceID is a clustering algorithm for the identification of cell types from single-cell RNA-sequencing data. It was specifically designed for the detection of rare cells which correspond to outliers in conventional clustering methods.

This module performs filtering, normalisation, and batch effect removal in the same step.

Example Usage: Inspecting the Aggregated Expression for a Group of Genes

Our cells come from 5 different batches (I5,II5,III5,IV5,V5) and are labelled to reflect this (i.e. "I5_1", "I5_2", ..., "I5_129", "II5_1", ..., "V5_236" )

We wish to filter out the gene Lpca5 and Atk2 which we know in advance will saturate our analysis with unwanted expression.

We will also be interested in the cluster that contains significant expression for Apoa genes (Apoa1, Apoa1bp, Apoa2, Apoa4, Apoa5).

First, we must load in our count matrix in order to correct for batch effects, filter out unwanted genes, and compute our clusters and outliers.

  • Mode of AnalysisCluster
    • Count Matrix → [input tabular]
    • Filtering:
      • Use Defaults?No
      • Batch Regex → "^I5,^II5,^III5,^IV5,^V5"
      • FGenes → "Lpca5,Atk2"

A PDF report will be generated giving metrics about the library size and number of features as histograms, and additional metrics relating to cell-cycle correction will be produced if that option has been selected.

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