thank you Jimmy. 
I will try the search w/o refinement. 
I will let you know if the problem solved. 

SunSun

On Monday, October 6, 2014 1:13:01 AM UTC+8, Jimmy Eng wrote:
>
> I don't know what the issue is with X!Tandem but just wanted to comment on 
> an unrelated issue which is that you should think twice about using 
> X!Tandem's refinement mode, especially if you are going to push those 
> search results through PeptideProphet or even simple target-decoy FDR.  All 
> proteins that make it to the refinement step (and their corresponding 
> peptides identified before or after refinement) will all have 
> distorted/high statistics.  Here are two publications that discuss this 
> problem:
>
> http://www.ncbi.nlm.nih.gov/pubmed/19947654
> http://www.ncbi.nlm.nih.gov/pubmed/21288048
>
> The simple solution is to not use refinement mode.
>
> On Sun, Oct 5, 2014 at 1:09 AM, zeyu sun <[email protected] <javascript:>> 
> wrote:
>
>> Hi, Dear SPC users and developers:
>> I'm wondering if anyone have some clue or remedy to the problem I have 
>> with the X!tandem search in TPP(v4.7.1):
>> I have some Orbitrap Velos data in RAW and converted them to mzXML files 
>> by MSconverter (ProteoWizard). 
>> I ran the Comet pipeline on them and everything was just fine. 
>> However, when I search the same mzXMLs by Tandem, some of those files 
>> gave error reports:
>> _________________________________________
>> X! TANDEM Jackhammer TPP (2013.06.15.1 - LabKey, Insilicos, ISB) 
>>
>> Loading spectra (mzParser)............................ loaded.
>> Spectra matching criteria = 55267
>> Pluggable scoring enabled.
>> Starting threads . started.
>> Computing models:
>>     12345678
>> *********
>> Warning: One or more sequences appear to contain invalid amino acid codes.
>>
>> *********
>>
>> 9    sequences modelled = 9 ks
>> Model refinement:
>>     partial cleavage .......... done.
>>     finishing refinement ... done.
>> Creating report:
>>     initial calculations  ..... done.
>>     sorting  ..... done.
>>     finding repeats ..... done.
>>     evaluating results ..... done.
>>     calculating expectations ..... done.
>>     writing results ..... done.
>>
>> command "c:\Inetpub\tpp-bin\tandem 
>> c:/Inetpub/wwwroot/ISB/data/xxx.tandem.params" failed: Unknown error
>>
>> Command FAILED
>> RETURN CODE:65280
>>  _________________________________________
>> Despite this error message, the .tandem files were generated, 
>> When I use the command tandem2XML to convert the .tandem files to 
>> .pep.xml, as expected, it didn't work out:
>> Operation not permitted
>> Command FAILED
>>  RETURN CODE:256
>>
>> It only happens in few mzXML files, not all of them. And Comet runs OK on 
>> all files, so I guess the mzXMLs should be fine. 
>> I further checked .tandem files, looks like they contain all necessary 
>> parts of Xtandem search results, and all spectra-peptide matches were 
>> listed there with scores. I just don't know why such files still gave error 
>> message and cannot go through tandem2XML. 
>> Please help! 
>> Thank you very much!
>>
>> SunSun
>>
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>

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