source: trunk/se/lu/thep/affymetrix/README @ 276

$Id: README 276 2007-05-21 17:44:04Z peter $

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{{{
Copyright (C) 2006 Jari Häkkinen
Copyright (C) 2007 Peter Johansson

This file is part of se.lu.thep.affymetrix package for BASE,
http://baseplugins.thep.lu.se/wiki/se.thep.lu.se.affymetrix

se.lu.thep.affymetrix package for BASE is free software; you can
redistribute it and/or modify it under the terms of the GNU General
Public License as published by the Free Software Foundation; either
version 2 of the License, or (at your option) any later version.

se.lu.thep.affymetrix package for BASE is distributed in the hope that
it will be useful, but WITHOUT ANY WARRANTY; without even the implied
warranty of MERCHANTABILITY or FITNESS FOR A PARTICULAR PURPOSE. See
the GNU General Public License for more details.

You should have received a copy of the GNU General Public License
along with this program; if not, write to the Free Software
Foundation, Inc., 59 Temple Place - Suite 330, Boston, MA 02111-1307,
USA.
}}}
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= About se.lu.thep.affymetrix package =

Send comments, suggestion, complaints, or questions about this
document to jari@thep.lu.se

This file contains information on se.lu.thep.affymetrix package. This
package contain the RMAExpress and Plier plug-ins.

The RMAExpress plug-in is installed in the BASE2 demo server
(http://lev.thep.lu.se/base2demo).


For installation or updating instructions please see file INSTALL.


What is RMAExpress and Plier plug-ins for BASE?

RMAExpress and Plier are two similar plug-ins needed to make use of
Affymetrix data in BASE. Affymetrix data can be imported into BASE as
files, but if one wants to perform clustering and other analysis
available in BASE, the data in these files have to be extracted and
imported into BASE.

These plug-ins do not implement the RMA method and Plier method,
respectively, but rather relies on third-party binary packages.

The RMAExpress plug-in relies on RMAExpressConsole available in the
RMAExpress package available at http://rmaexpress.bmbolstad.com/. The
RMA algorithm is published in scientific journals, for more
information please refer to the RMAExpress web site. In short RMA is
the Robust Multichip Average. It consists of three steps: a background
adjustment, quantile normalization, and finally a summarization.

The Plier plug-in relies on `apt-probeset-summarize` included in
Affymetrix Power Tools (APT) available at
http://www.affymetrix.com/support/developer/powertools/index.affx.
`apt-probeset-summarize` provides several methods (see Affymetrix home
page). Here we use the method given with option flag `--option
plier-mm`.

What is required to use the these plug-ins for BASE? The "BASE
getting started document" may also be helpful,
http://base.thep.lu.se/chrome/site/doc/user/getting_started.html

1) You need to install the plug-in and make BASE aware of the new
   plugin, see INSTALL for instruction for installation
   instructions.

2) You need to import probe set information into BASE (reporter
   information), i.e. import AffyChip_annot.csv into BASE. This is
   only needed to do once, and optionally later if reporter
   information should be updated. This step can only be performed by a
   user with reporter administrative (create) privileges, an ordinary
   BASE user does NOT have this privilege.

    - First you need to create an plug-in configuration for the
      'Reporter importer' plug-in. Locate the 'Reporter importer'
      plug-in definition ('Administrate' -> 'Plugins' ->
      'Definitions'). Click on it and choose 'New configuration ...',
      name the configuration and choose 'Save and configure'.

    - A 'Parser settings' dialog will appear. Set fields according to
      this list:

  Data header  : "Probe Set ID","GeneChip Array",.*
  Data splitter: (?!"),(?=")
  Remove quotes: true
  Name         : \Probe Set ID\
  Reporter ID  : \Probe Set ID\

      Finalize by clicking 'Next'.

    - Import the reporters by choosing 'View' -> 'Reporters' and the
      clicking on 'Import...'. Click 'Next' without changing the 'auto
      detect' settings. Supply the file name in the next dialog, click
      'Next'. Set the 'Update existing reporters' to "true" if an
      update of already stored reporter information is wanted. Start
      the import by clicking 'Next' in the parameter dialog

   The reporter information is now in the database. You may delete the
   AffyChip_annot.csv file from BASE if you wish, it is not needed
   anymore. You may want to update reporter information when
   Affymetrix releases a new version of their annotation but remember
   there is no versioning for this type of information in BASE. If you
   update reporter information you only need to redo the last step.

3) You need to define the design of the AffyChip in BASE, i.e. import
   the AffyChip.cdf file. This is only needed to do once for every
   AffyChip design to be added into BASE. This step can only be
   performed by a user with array LIMS administrative (create)
   privileges, an ordinary BASE user does NOT have this privilege.

    - Define a new array design through 'Array LIMS' -> 'Array
      Designs'. Click on 'New ...'. In the 'Create array design'
      dialog, set the 'Name' (use the filename of the CDF file for
      convenience), tick the Affy chip tick box, choose a CDF file,
      and optionally write a description. Click on next, and the new
      design is created.

    - Remember to share the design to all users/roles/groups that
      should have access to it (sharing it to group Everyone should be
      sufficient in most cases).

4) Create an experiment, i.e., grouping .cel files into an experiment.

    - Upload the .cel files into BASE.

    - Create raw bio assay for these files, one per file: Choose
      'View' -> 'Raw bioassays', click on 'New ...'. In the dialog set
      the 'Name', choose the proper 'Raw data type' (i.e.,
      "Affymetrix"), choose the 'Array design' that matches the .cel
      file, and choose the 'CEL file'. (The other fields are not
      needed but may be handy to fill in.) Click on 'Save'.

    - Create the experiment: Choose 'View' -> 'Experiment', click on
      'New ...'. In the dialog set the 'Name', choose the proper 'Raw
      data type' (i.e., "Affymetrix"), and add 'Raw bioassays'.

5) Select the experiment and create a new root bioassay set for it.

    - Locate the newly created experiment and click on 'Analyze'.

    - Click on 'New root bioassay set ...' and click 'Next' in the
      pop-up dialog. Select the 'Raw bioassays' to include in the new
      root bioassay set and type a name for the bioassay set. Click on
      'Next' to start the plug-in.

6) When the plug-in is finished the resulting expression values will
   be accessible in BASE.