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The ChIP-seq pipeline “Cistrome” integrated into UGENE allows one to do the following analysis steps: peak calling and annotating, motif search and gene ontology. ChIP-seq analysis is started from MACS tool. CEAS then takes peak regions and signal wiggle file to check which chromosome is enriched with binding/modification sites, whether bindings events are significant at gene features like promoters, gene bodies, exons, introns or UTRs, and the signal aggregation at gene transcription start/end sites or meta-gene bodies (average all genes). Then peaks are investigated in these ways:

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Note that it is originally based on the General ChIP-seq pipeline from the public Cistrome installation on the Galaxy workflow platform.

Environment requirements:

Before proceeding, please make sure that your computer is configured as follows: 

  1. The following software packages are installed:

  2. Make sure R and Python are added to the PATH environment variable.

  3. Move directory “cistrome” from the Cistrome Configuration Package, that goes with this tutorial, to the “data” subdirectory of the UGENE installation directory.

  4. Install required Bioconductor packages. To do it, please make sure your computer is connected to the internet, unpack “installRpackages.R” script from the Cistrome Configuration Package and run it as follows:

  • On Linux/Mac OS X open a terminal and run commands: 
    •  “sudo Rscript –vanilla installRpackages.R”
  • It may be required to also run the following command on Linux: 
    •  “sudo apt-get install libxml2-dev”
  • On Windows search for “cmd” executable file, select “Run as administrator” item in its context menu and run command: 
    •  “Rscript –vanilla installRpackages.R”

Note that it takes some time to install the packages.

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Download and install the UGENE NGS package to use this pipeline.

Select Samples tab on the Workflow Designer Palette and double-click on the ChIP-seq analysis with Cistrome tools sample. The following configure wizard appears:

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Output directory

Directory to store seqpos results.

Output file name

Name of the output file which stores new motifs found during a de novo search.

 

Peak2Gene output:

Gene annotations

Location of peak2gene gene annotations data file..

Peak annotations

Location of peak2gene peak annotations data file..

 

Conduct GO output:

 

Output directory

Directory to store Conduct GO results.

Choose these output directories click on the Finish button. 

 Note that default button reverts all parameters to default settings. 

 Now let’s validate and run the workflow. To validate that the workflow is correct and all parameters are set properly click the Validate workflow button on the Workflow Designer toolbar:

 

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If there are some errors, they will be shown in the Error list at the bottom of the Workflow Designer window, for example:

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  <img src="/wiki/download/attachments/32446173244619/ChIP-seq analysis with Cistrome tools_12.png"/>
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The dashboard will contain information about workflow: input and output files, all information about task.  . 

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The work on the Cistrome pipeline was supported by grant RUB1-31097-NO-12 from NIAID.