Gene-level RNA-Seq Data Analysis (lagacy)
  • Introduction
  • RNA-Seq Analysis Workflow
    • Login to server
    • Obtain data and software
    • Create mapping indices
    • Mapping with STAR
    • Quantification using RSEM
  • De novo assembly using Trinity
    • De novo assembly of RNA-Seq reads
    • Compare de novo reconstructed transcripts to reference annotations
    • Quantification using RSEM
  • Differential expression analysis using R
    • Install R libraries
    • Perform DE analysis
    • Perform DE analysis (Trinity)
    • Extracting DE transcripts and generating heatmaps (Trinity)
  • Visualization
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  • Use biocLite()
  • Install from source

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  1. Differential expression analysis using R

Install R libraries

The latest R version is 3.3.2 at the time of writing. Type R to start the R environment.

R version 3.3.2 (2016-10-31) -- "Sincere Pumpkin Patch"
Copyright (C) 2016 The R Foundation for Statistical Computing
Platform: x86_64-pc-linux-gnu (64-bit)

R is free software and comes with ABSOLUTELY NO WARRANTY.
You are welcome to redistribute it under certain conditions.
Type 'license()' or 'licence()' for distribution details.

  Natural language support but running in an English locale

R is a collaborative project with many contributors.
Type 'contributors()' for more information and
'citation()' on how to cite R or R packages in publications.

Type 'demo()' for some demos, 'help()' for on-line help, or
'help.start()' for an HTML browser interface to help.
Type 'q()' to quit R.

>

Within the R console, enter following to initiate Bioconductor:

source("http://bioconductor.org/biocLite.R")
chooseBioCmirror(ind=6)     # Choose "Riken, Kobe (Japan)"

Use biocLite()

R will ask you if you wish to install these in your personal library in ~/R. Type "y" to both questions.

Would you like to use a personal library instead?  (y/n) y
Would you like to create a personal library
~/R/x86_64-unknown-linux-gnu-library/3.3
to install packages into?  (y/n) y

When completed, R will prompt you if you wish to update other packages. Choose "n" to skip this step to save time.

trying URL 'http://bioconductor.jp/packages/3.4/bioc/src/contrib/BiocInstaller_1.24.0.tar.gz'
Content type 'application/x-gzip' length 17756 bytes (17 Kb)
opened URL
==================================================
downloaded 17 Kb
...
...
...
The downloaded source packages are in
        ‘/tmp/RtmpAkPtJ1/downloaded_packages’
Old packages: 'BiocInstaller', 'boot', 'caret', 'class', 'cluster',
  'codetools', 'foreach', 'foreign', 'gdata', 'GenomeInfoDb', 'gplots',
  'gridSVG', 'gtools', 'httpuv', 'iterators', 'KernSmooth', 'lattice', 'lme4',
  'MASS', 'Matrix', 'matrixStats', 'mgcv', 'mime', 'nlme', 'nnet', 'plyr',
  'quantreg', 'R6', 'Rcpp', 'RcppEigen', 'RCurl', 'rpart', 'scales', 'shiny',
  'SparseM', 'spatial', 'stringr', 'survival', 'XML', 'xtable'
Update all/some/none? [a/s/n]: n

Install from source

To install limma, run the following command in R:

install.packages("https://bioconductor.org/packages/release/bioc/src/contrib/limma_3.30.2.tar.gz")

Install limma v3.30.2 without error:

...
...
* installing *source* package ‘limma’ ...
...
...

** R
** inst
** preparing package for lazy loading
** help
*** installing help indices
** building package indices
** installing vignettes
** testing if installed package can be loaded
* DONE (limma)

To install edgeR, run the following command in R:

install.packages("https://bioconductor.org/packages/release/bioc/src/contrib/edgeR_3.16.1.tar.gz")

Install edgeR v3.16.1 without error:

...
...
* installing *source* package ‘edgeR’ ...
...
...

** R
** inst
** preparing package for lazy loading
** help
*** installing help indices
** building package indices
** installing vignettes
** testing if installed package can be loaded
* DONE (edgeR)

Use sessionInfo() to print version information about R and attached or loaded packages. This information is useful when providing bug report to package developers.

R version 3.3.2 (2016-10-31)
Platform: x86_64-unknown-linux-gnu (64-bit)

locale:
 [1] LC_CTYPE=en_US.UTF-8       LC_NUMERIC=C
 [3] LC_TIME=en_US.UTF-8        LC_COLLATE=en_US.UTF-8
 [5] LC_MONETARY=en_US.UTF-8    LC_MESSAGES=en_US.UTF-8
 [7] LC_PAPER=en_US.UTF-8       LC_NAME=C
 [9] LC_ADDRESS=C               LC_TELEPHONE=C
[11] LC_MEASUREMENT=en_US.UTF-8 LC_IDENTIFICATION=C

attached base packages:
[1] stats     graphics  grDevices utils     datasets  methods   base

other attached packages:
[1] BiocInstaller_1.24.0

Enter q() to exit the R console. Enter "n" to not save the workspace as an image.

PreviousDifferential expression analysis using RNextPerform DE analysis

Last updated 5 years ago

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To install or update Bioconductor packages and , and CRAN package enters biocLite(c("cluster","Biobase","qvalue"))

You can usually use biocLite() to install Bioconductor packages. But, if R version you are using is out-dated, the packages that can be installed using biocLite() will also be of older versions. Therefore, in this case, you may choose to install the packages from source. Below I showed how you can install and from package sources from .

Biobase
qvalue
cluster
limma
edgeR
http://bioconductor.org/