Luis,

Thanks for your message. I rerun the experiment leaving all options same
except -he option [-he evalcut command line option to omssacl] for OMSSA
seach set to 1,000,000 then

I got total hits of 309 while running ProphetModels.pl corresponding to 575
decoys and 1478 non-decoys. Here is the brief program output ....

----------------------------------------------------------------------------------------------------------

read in 187 1+, 885 2+, 981 3+, 0 4+, 0 5+, 0 6+, and 0 7+ spectra.

Found 575 Decoys, and 1478 Non-Decoys

--------------------------------------------------------------------------------------------------------------


Here is the output from the corresponding previous run with -he=1

read in 0 1+, 99 2+, 31 3+, 0 4+, 0 5+, 0 6+, and 0 7+ spectra.
Initialising statistical models ...
Found 0 Decoys, and 130 Non-Decoys

----------------------------------------------------------------------------------------------------


In the current case study I have used the following commandline options
while running OMSSA search ....

---------------------------------------------------------------------------------------------------------------------------------------------------------------
/home/APCF/omssa-2.1.7.linux/omssacl  -cl 0 -d
/mnt/sanfs/APCF/databases/SwissProt/decoy_uniprot_sprot_generic.fasta -e 0
-fm /home/APCF/omssa/spectra/0f3ad159ab3ed187e2d43fe8d6134dac_3.mgf -he
1000000 -hl 10 -ht 6 -i 1,4 -mm 128 -mux
/home/APCF/omssa-2.1.7.linux/usermods.xml -mx
/home/APCF/omssa-2.1.7.linux/mods.xml -ox
/home/APCF/omssa/results/0f3ad159ab3ed187e2d43fe8d6134dac/0f3ad159ab3ed187e2d43fe8d6134dac_3.omx
-pc 1 -sct 0 -sp 100 -te 2 -tem 0 -to 0.8 -tom 0 -v 1 -w  -x 0 -z1 0.95 -zh
3 -zl 1 -zt 3
---------------------------------------------------------------------------------------------------------------------------------------------------------------

After OMSSA search, I use omssa2pepXML command from omssa distribution
(omssa-2.1.7.linux) to generate pepXML file.

When I run ProphetModels.pl after InterProphetParser merger then I see the
following ....
-----------------------------------------------------------

Parsing search results "/mnt/sanfs/APCF/results/tpp/2010-09-08/jagan-J1557
(MASCOT)"...

=> Total of 119 hits.

Parsing search results "jagan-J1558 (X! Tandem (k-score))"...

=> Total of 239 hits.

Parsing search results "/mnt/sanfs/APCF/results/tpp/2010-09-08/jagan-J1561
(OMSSA)"...

=> Total of 152 hits.

----------------------------------------------------------------
here are the similar entries corresponding to previous run where the OMSSA
command line option used for -he was 1

----------------------------------------------------------------------------------------------------------------------------------

Parsing search results "/mnt/sanfs/APCF/results/tpp/2010-09-03/jagan-J1366
(MASCOT)"...

=> Total of 124 hits.

Parsing search results "jagan-J1367 (X! Tandem (k-score))"...

=> Total of 335 hits.

Parsing search results "/mnt/sanfs/APCF/results/tpp/2010-09-03/jagan-J1370
(OMSSA)"...

=> Total of 0 hits.

-------------------------------------------------------------
The results are so much different could you mind to advise which is the
better value for OMSSA command line option -he to choose.

with regards,

Jagan Kommineni



On Tue, Sep 7, 2010 at 11:16 AM, Luis Mendoza
<[email protected]>wrote:

> Hello Jagan,
> Based on the output you provided, it seems that you are not giving
> PeptideProphet that many search results to begin with:
>
>  read in 0 1+, 99 2+, 31 3+, 0 4+, 0 5+, 0 6+, and 0 7+ spectra.
>
> Which corresponds to the 130 reported results.  How did you search your
> data through OMSSA?  How was the input file jagan-J1370.pep.xml  generated?
>  How many results do you see when you open this file in PepXMLViewer,
> especially compared to the Mascot and Tandem pep.xml results?  Ultimately,
> the model fails due to the very low number of search hits in the input file.
>
> We will look into providing a better error message when this is
> encountered, since "exit code: 11" is not too useful!
> --Luis
>
>
>
> On Mon, Sep 6, 2010 at 5:55 PM, Jagan Kommineni <[email protected]
> > wrote:
>
>>
>>
>> On Fri, Sep 3, 2010 at 2:51 PM, Jagan Kommineni <
>> [email protected]> wrote:
>>
>>> Dear All,
>>>
>>> I have taken RAW file generated by Instrument info: Manufacturer: Thermo
>>> Finnigan, Model: LCQ Deca, Ionization: nanoelectrospray, Analyzer:
>>> quadrupole ion trap, Detector: electron multiplier and run standard search
>>> using Mascot, X!tandem and OMSSA and fed the data through TPP pipeline then
>>> I found that 0 (zero) decoy and 130 non-decoy entries for the search
>>> results of OMSSA whereas 545 Decoys and 785 Non-Decoys entries for
>>> Mascot search results and 536 Decoys and 833 Non-Decoys entries for
>>> x!tandem search.
>>>
>>> I found total hits retruned from Mascot and X!tandem search result sets
>>> respectively are of 303 and 351.
>>>
>>>
>>> For OMSSA based search results, the PeptideProphetParser program quitted
>>> with exit code of 11.
>>>
>>> Here is the complete STDOUT of OMSSA and TPP pipeleine ...
>>> ----------------------------------------------------------------------------------------------------------------------------------------------------------------------------------
>>>
>>> /home/APCF/tpp/bin/xinteract -Njagan-J1370.pepprophet.xml -eT
>>> -d'sp|decoy' -p0.05 -l7 -x20 -R -OP jagan-J1370.pep.xml
>>>
>>> /home/APCF/tpp/bin/xinteract (TPP v4.4 JETSTREAM (unstable development
>>> prerelease) rev 0, Build 201007011135 (linux))
>>>
>>> running: "/mnt/sanfs/APCF/APCF_WEB/tpp/bin/InteractParser
>>> 'jagan-J1370.pepprophet.xml' 'jagan-J1370.pep.xml' -P -L'7' -E'trypsin'"
>>>  file 1: jagan-J1370.pep.xml
>>>  processed altogether 130 results
>>>
>>>
>>>  results written to file
>>> /mnt/sanfs/APCF/results/tpp/2010-09-03/jagan-J1370.pepprophet.shtml
>>>
>>>
>>>
>>> command completed in 0 sec
>>>
>>> running: "/mnt/sanfs/APCF/APCF_WEB/tpp/bin/PeptideProphetParser
>>> 'jagan-J1370.pepprophet.xml' DECOY='sp|decoy' MINPROB=0.05 EXTRAITRS=20
>>> NONPARAM"
>>> Using Decoy Label "sp|decoy".
>>> Using non-parametric distributions
>>>  (OMSSA)
>>> WARNING!! The discriminant function for OMSSA is not yet complete.  It is
>>> presented here to help facilitate trial and discussion.  Reliance on this
>>> code for publishable scientific results is not recommended.
>>> init with OMSSA trypsin
>>> MS Instrument info: Manufacturer: UNKNOWN, Model: UNKNOWN, Ionization:
>>> UNKNOWN, Analyzer: UNKNOWN, Detector: UNKNOWN
>>>
>>>  PeptideProphet  (TPP v4.4 JETSTREAM (unstable development prerelease)
>>> rev 0, Build 201007011135 (linux)) akel...@isb
>>>  read in 0 1+, 99 2+, 31 3+, 0 4+, 0 5+, 0 6+, and 0 7+ spectra.
>>> Initialising statistical models ...
>>> Found 0 Decoys, and 130 Non-Decoys
>>> WARNING: No decoys with label sp|decoy were found in this dataset.
>>> reverting to fully unsupervised method.
>>> Iterations: .........10.........20
>>>
>>> command "/mnt/sanfs/APCF/APCF_WEB/tpp/bin/PeptideProphetParser
>>> 'jagan-J1370.pepprophet.xml' DECOY='sp|decoy' MINPROB=0.05 EXTRAITRS=20
>>> NONPARAM" exited with non-zero exit code: 11
>>> QUIT - the job is incomplete
>>>
>>> ==============================================================================================
>>>
>>> I wonder is there any requirement to have at least few decoy entries for
>>> PeptideProphetParser to continue without fail?
>>>
>>> Further study with pep XML generated by different algorithm search
>>> results, I found that there is only one decoy entry for xml tag of
>>> search_hit which I am here with enclosing in this mail ...
>>>
>>> ------------------------------------------------------------
>>> <search_hit hit_rank="2" peptide="LGDSEVDIGIMANLDIVESKR"
>>> peptide_prev_aa="K" peptide_next_aa="L" protein="520685"
>>> num_tot_proteins="2" num_matched_ions="21" tot_num_ions="40"
>>> calc_neutral_pep_mass="2273.156" massdiff="0.041999999999865"
>>> is_rejected="0" protein_descr="sp|decoy_C1ERL6|ACDH_BACC3 Reverse sequence,
>>> was Acetaldehyde dehydrogenase OS=Bacillus cereus (strain 03BB102)
>>> GN=BCA_2154 PE=3 SV=1">
>>>           <alternative_protein protein="520683"
>>> protein_descr="sp|decoy_A0RD69|ACDH_BACAH Reverse sequence, was Acetaldehyde
>>> dehydrogenase OS=Bacillus thuringiensis (strain Al Hakam) GN=BALH_1843 PE=3
>>> SV=1"/>
>>>           <search_score name="pvalue" value="0.000012588862528"/>
>>>           <search_score name="expect" value="0.648238298133515"/>
>>>         </search_hit>
>>> -----------------------------------------------------------
>>>
>>> whereas I found 581 and 572 similar tags in Mascot and X!tandem based
>>> results,
>>>
>>> with regards,
>>>
>>> --
>>> Dr. Jagan Kommineni
>>> Ludwig Institute for Cancer research
>>> Pakville VIC 3145
>>> Australia.
>>>
>>>
>>
>>
>> --
>> Dr. Jagan Kommineni
>> Ludwig Institute for Cancer research
>> Pakville VIC 3145
>> Australia.
>>
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-- 
Dr. Jagan Kommineni
Ludwig Institute for Cancer research
Pakville VIC 3145
Australia.

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