Hi SunSun,

I ran a search today and got the same error.  I was using a bovine database 
that I downloaded quite a while ago.  So I downloaded the latest version today, 
appended the CRAP database to it and then ran it through the decoy utility to 
create a concatenated forward and reversed sequence database.  Subsequent 
searches with the new database completed without error.

I can only presume the older database became corrupted somehow.  I don't know 
of a utility that will validate the integrity of a database.  If you know regex 
or maybe perl you could write a script to look for characters that are not part 
of the amino acid code.  For me the expedient solution was to download a fresh 
version of the database.

Brian

> On Oct 6, 2014, at 12:35 PM, zeyu sun <[email protected]> wrote:
> 
> Thank you Eric, 
> Is there any tool out there to check the validity of my fasta?
> 
> To Jimmy:
> I just tried the tandem search without refinement, the problem is still 
> there. 
> 
> Thank you all for the reply! 
> 
> SunSun 
> 
>> On Monday, October 6, 2014 10:58:12 PM UTC+8, Eric Deutsch wrote:
>> I think the key part in your output is this:
>> 
>>  
>> 
>> *********
>> Warning: One or more sequences appear to contain invalid amino acid codes.
>> 
>> *********
>> 
>> 
>> I think this is a warning that we put in to warn that X!Tandem may crash 
>> because of your database. You should check your database for validity. 
>> Perhaps there are some strange characters in one of the sequences that 
>> causes X!Tandem to crash sometimes, but not Comet. We have seen this before.
>> 
>>  
>> 
>> Eric
>> 
>>  
>> 
>>  
>> 
>>  
>> 
>> From: [email protected] [mailto:[email protected]] On 
>> Behalf Of zeyu sun
>> Sent: Monday, October 06, 2014 7:45 AM
>> To: [email protected]
>> Subject: Re: [spctools-discuss] X!tandem error
>> 
>>  
>> 
>> 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]> 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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