How to download and use ReAdW
IonSource.com

Implementing the ISB program ReAdW for the conversion of Xcalibur raw files to the new universal mzXML format. Your new mzXML file can then be viewed with the free Insilicos viewer and searched on the public version of the Phenyx .

(Even though this software download did not cause us any trouble our policy is to  never download software from the internet onto a "mission critical computer."  Also, the software described here is independent of IonSource so please read our terms of use disclaimer and theirs)

 

Download  the latest version of "ReAdW" , also you can view some additional ReAdW info by following this link.  Thank the Institute for Systems Biology (the ISB) for this software.  ReAdW requires that you have Xcalibur installed on your computer. SourceForge does supply an additional tool if you do not have access to Xcalibur.
Place the ReAdW program in the same folder as the Xcalibur raw data file, see Figure 1, bottom.  You will need to run ReAdW from the "RUN" dialog see top gif of Figure 1.  ReAdW is easy to download and to run.  Download ReAdW to the folder that contains your data file.

1.) Go to the START menu on your computer, and click on the "Run..." tab. 
2.) Click on RUN, a dialog box will come up, type cmd  
3.) Type cd\
4.) Type cd "your path to the folder"  mzXML is the folder in this example.
5.) Type, ReAdW yourrawfilename.raw c, as shown in Figure 1 and hit return.  The "c" is for centriod data.
6.) Wait until the converting process is finished.

 

Here is a screen shot of the run dialog and the folder.  Notice in the folder view that the created mzXML file is 1.4 times bigger.

Figure 1
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You can  type "program name /help" to get program usage information as shown above.


To run the program type "ReAdW datafile name c" and hit return and wait until you see "done" as shown above.


Download the free Insilicos Viewer at http://www.insilicos.com/products.html

Figure 2

The free viewer comes with an installer and installs painlessly.  It is a small program and very efficient so it launches quickly and also opens files quickly.

Once the viewer is installed open the program and open the file you just converted to mzXML format.  It is as easy as that!  It is a revolution I think.  Now anyone can read native Finnigan Xcalibur files.  Thank you Insilicos and ISB!  Here are a few screen shots of the program.

Figure 3

Figure 3: This is a screen shot of the viewer with a base peak plot in the upper left hand frame and a full MS spectrum in the bottom plot.  Notice the tabs at the bottom, the first tab is the full MS followed by the three MS2s that were performed.

Figure 4

Figure 4: Shows a screen shot of the viewer with a base peak plot in the upper left hand frame and an MS2 spectrum in the bottom plot. 

Now that you have your data in the new common file format, mzXML, you can submit your files to those proteoimc search engines that accept mzXML.  If your vendor does not support mzXML tell them that you want it. 

One of the search engines that accepts mzXML is Phenyx from GeneBio.  You can download the Public Phenyx  and submit your new mzXML file for protein database searching.  Follow this link to the Phenyx public search engine.

 

Figure 5
This is the Public Phenyx data submission page.  Browse to your new mzXML file to begin your search.

 

Below is the second part of the submission page. Notice that there is an option for a second round search in which you can include more variables.

 

 

Figure 6


This is the Phenyx Results Page.  Look at how well the horse myoglobin hit differentiates itself from the second hit, scores 108.87 and 31.0 respectively.   Pretty amazing!  That OLAV scoring must be doing something  right!  The protein coverage is 66%.  Click on the details link to see the protein details page.  If you would like to do your own Phenyx search click here to download the mzXML file.

 

 

This is the protein match details page.  All of the peptides identified are listed here as well as the protein sequence coverage map.

 

Shown below is the sequence coverage map.  Notice the half cleaved peptides and those matches that were deemed invalid.

 

Figure 7
Below is the peptide match details page.  Remember this was all from an mzXML file generated from a native Xcalibur raw file.  Also remember that you can download tools from SourceForge and ISB that can convert other MS formats to mzXML!   We have also noticed that Phenyx does a very good job at identifying, scoring, and correlating difficult spectra.

 

Conclusion:  A common file format has been a dream of mass spectrometrists since the second instrument company made a mass spectrometer.  mzXML has advanced that dream, it is our responsibility to encourage vendors to support this format.  Thanks to ReAdW.exe we can now give our clients native data files and point them to ISB or Insilicos to get the converter and the viewer.  I have every confidence that this type of open source effort will speed up scientific research.  "Enable the researcher" should be our motto.  Some search engine sellers or MS vendors may fear this advance, but what it will really mean is that more people will have access to MS data and more people will be able to search their own data, and hence more search engine software will be sold.  It will be good for everyone.
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Last updated:  Tuesday, January 19, 2016 02:51:41 PM

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

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