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SARTools

SARTools is a R package dedicated to the differential analysis of RNA-seq data. It provides tools to generate descriptive and diagnostic graphs, to run the differential analysis with one of the well known DESeq2 or edgeR packages and to export the results into easily readable tab-delimited files. It also facilitates the generation of a HTML report which displays all the figures produced, explains the statistical methods and gives the results of the differential analysis. Note that SARTools does not intend to replace DESeq2 or edgeR: it simply provides an environment to go with them. For more details about the methodology behind DESeq2 or edgeR, the user should read their documentations and papers.

SARTools is distributed with two R script templates (template_script_DESeq2.r and template_script_edgeR.r) which use functions of the package. For a more fluid analysis and to avoid possible bugs when creating the final HTML report, the user is encouraged to use them rather than writing a new script. Two other scripts are available (template_script_DESeq2_CL.r and template_script_edgeR_CL.r) to run SARTools in a shell with the Rscript command. In that case, the optparse R package must be available to interpret the command line parameters.

How to install SARTools?

Within R

In addition to the SARTools package itself, the workflow requires the installation of several packages: DESeq2, edgeR, genefilter, xtable and knitr (all available online, see the dedicated webpages). SARTools needs R version 3.3.0 or higher, DESeq2 1.12.0 or higher and edgeR 3.12.0 or higher: old versions of DESeq2 or edgeR may be incompatible with SARTools.

To install the SARTools package from GitHub, open a R session and:

  • install DESeq2, edgeR and genefilter with source("http://bioconductor.org/biocLite.R") and biocLite(c("DESeq2", "edgeR", "genefilter")) (if not installed yet)

  • install devtools with install.packages("devtools") (if not installed yet)

  • Notes:

    • Ubuntu users may have to install some libraries (libxml2-dev, libcurl4-openssl-dev and libssl-dev) to be able to install DESeq2 and devtools
    • Some users may have to install the pandoc and pandoc-citeproc libraries to be able to generate the final HTML reports
  • for Windows users only, install Rtools or check that it is already installed (needed to build the package)

  • load the devtools R package with library(devtools)

  • run install_github("PF2-pasteur-fr/SARTools", build_vignettes=TRUE)

Using Conda

  1. Install miniconda2
  2. Install the SARTools R library and its dependencies using conda conda install r-sartools

Note: if you want to set a dedicated conda environment for SARTools, use conda create -n sartools r-sartools and follow the instructions to activate it.

How to use SARTools?

A HTML vignette is available within the vignettes folder on GitHub and provides extensive information on the use of SARTools. The user can also open it with vignette("SARTools") if it has been generated during the installation of the package.

Be careful to use the R script associated with the version of SARTools installed on your system.

Please read the NEWS file to see the latest improvements!

About SARTools

The SARTools package has been developped at PF2 - Institut Pasteur by M.-A. Dillies and H. Varet (hugo.varet@pasteur.fr). Thanks to cite H. Varet, L. Brillet-Guéguen, J.-Y. Coppee and M.-A. Dillies, SARTools: A DESeq2- and EdgeR-Based R Pipeline for Comprehensive Differential Analysis of RNA-Seq Data, PLoS One, 2016, doi: http://dx.doi.org/10.1371/journal.pone.0157022 when using this tool for any analysis published.

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Version

Version

1.6.2

License

GPL-2

Maintainer

Hugo Varet

Last Published

June 1st, 2018

Functions in SARTools (1.6.2)

BCVPlot

BCV plot (for edgeR dispersions)
SARTools-package

Statistical Analysis of RNA-Seq Tools
cairoSizeWrapper

Cairo Size Wrapper
MAPlot

MA-plots
checkParameters.edgeR

Check the parameters (when using edgeR)
SERE

Pairwise SERE for two samples
checkParameters.DESeq2

Check the parameters (when using DESeq2)
clusterPlot

Clustering of the samples
pairwiseScatterPlots

Scatter plots for pairwise comparaisons of log counts
rawpHist

Histograms of raw p-values
dispersionsPlot

Plots about DESeq2 dispersions
descriptionPlots

Description plots of the counts
exploreCounts

Explore counts structure
writeReport.edgeR

Write HTML report for edgeR analyses
diagSizeFactorsPlots

Assess the estimations of the size factors
removeNull

Remove features with null counts in all samples
writeReport.DESeq2

Write HTML report for DESeq2 analyses
run.DESeq2

Wrapper to run DESeq2
MDSPlot

MDS plot (for edgeR objects)
PCAPlot

PCA of samples (if use of DESeq2)
countsBoxplots

Box-plots of (normalized) counts distribution per sample
densityPlot

Density plot of all samples
majSequences

Most expressed sequences per sample
barplotNull

Percentage of null counts per sample
nDiffTotal

Number of differentially expressed features per comparison
exportResults.DESeq2

Export results for DESeq2 analyses
barplotTotal

Total number of reads per sample
loadCountData

Load count files
exportResults.edgeR

Export results for edgeR analyses
loadTargetFile

Load target file
tabIndepFiltering

Table of the number of features discarded by the independent filtering (if use of DESeq2)
summarizeResults.edgeR

Summarize edgeR analysis
run.edgeR

Wrapper to run edgeR
summarizeResults.DESeq2

Summarize DESeq2 analysis
tabSERE

SERE statistics for several samples
volcanoPlot

Volcano plots