Hi,

Following a DOS batch file, which converts all *.mzXML data in a
directory to *.mgf, executes an OMSSA search with a decoy data base
and subsequently carries out the PeptideProphet/ProteinProphet
validation steps.

My raw data are generated on an Agilent XCTultra, which has the same
hardware and data format as the Bruker HCTultra. For conversion of
*.yep to *.mzXML I’m using the Bruker tool # compassxport -multi d:
\data\directory.

Note: I´m running a standard OMSSA search (binary *.oms output), for
comparison; usually the automated workflow with OMSSA/TPP delivers
more hits.

Of course, one might wish to change some parameters.

This file is copy & paste now. Unfortunately, I cannot upload the
*.bat.

---
omssavalidation.bat
---
@ECHO OFF
ECHO *** Converting mzXML files to mgf ***
FOR %%I IN (*.mzXML) DO MzXML2Search -mgf -ACID -M2-2 -P5 -I0.01 -B10 -
T5000.0 %%I

ECHO *** OMSSA search: Decoy data base, trypsin, pep.xml output ***
REM decoy entries in data base stdecoy start with decoy_; reverse
decoy data base generated with FastaTools 0.9 and merged with original
data base using mergeFasta.pl
FOR %%I IN (*.mgf) DO omssacl -d c:\blastdb\stdecoy -fm %%I -v 2 -mf 3
-mv 4 -he 1e6 -w -op %%I_OMSSA_TPP.pep.xml
FOR %%I IN (*.mgf) DO omssacl -d c:\blastdb\stdecoy -fm %%I -v 2 -mf 3
-mv 4 -w -ob %%I_OMSSA_DECOY.oms

ECHO *** Validation of OMSSA results with PeptideProphet/
ProteinProphet ***
FOR %%I IN (*OMSSA_TPP.pep.xml) DO interactparser ppval_%%I %%I -L5 -
Etrypsin -C -P
FOR %%I IN (ppval*.pep.xml) DO peptideprophetparser %%I DECOY=decoy
MINPROB=0 NONPARAM
FOR %%I IN (ppval*.pep.xml) DO refreshparser %%I c:\blastdb\stdecoy
FOR %%I IN (ppval*.pep.xml) DO proteinprophet %%I %%I.prot.xml
---

I´m using TPP v4.3r1, and OMSSA 2.1.7.

Currently I am testing this *.bat with several data files. It looks
promising, the *prot.shtml looks the same as with the built-in Tandem
workflow.

However, I cannot access directly the peptide mass spectra. Any ideas?

Comments and suggestions are welcome.

Hope, it might be useful.

Best regards,

Robert

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