Hi David.  Thanks for your response.

The problem was solved after I closely looked at the tandem.params file I
created after upgrading from 4.5.2 to 4.6.  I made a silly copy paste
error.  In the static modifications section I had two decimal points in one
of the modification mass specifications.  I am not certain how Tandem
treated this, but ASAPRatio treated it as multiple modifications to a
single residue, in this case it was [.

This was the erroneous modifications line:
<note type="input" label="residue, modification mass">57.021464@C
,48.085.047@[,48.085@K</note>

This is the correct modifications line:
<note type="input" label="residue, modification mass">57.021464@C,48.085@
[,48.085@K</note>

The Tandem API suggests that it will use the last instance of a
modification to the same residue e.g. 48@K,28@K then it will use 28@K and
ignore the 48@K.  However, in my case the mistake of 48.085.047@[ was
apparently treated as multiple modifications to [.  Useless anyway because
ASAPRatio could not do anything with that.  All the output I got was wrong
no matter how I specified the modification masses for ASAPRatio.  And
setting the -S static flag would cause ASAPRation to fail.

Brian


On Tue, Aug 28, 2012 at 7:29 PM, David Shteynberg <
[email protected]> wrote:

> Hi Brian,
>
> Have you tried removing the static option -S when running this command?
>
> Thanks,
> -David
>
> On Tue, Aug 28, 2012 at 2:50 PM, Brian Hampton <[email protected]>wrote:
>
>> Hello,
>>
>> I am trying the newest TPP installed on Win7 using the installer.  I have
>> two small data sets of differentially labelled peptides searched separately
>> with Tandem where one search specifies all light and the second all heavy
>> labelled modifications.  When running ASAPRatio, I select static
>> modifications and tell it N-terminus & K are modified.  It is pulling the
>> correct mods from the tandem result file. But I get "error: cannot have
>> variablel modified n in static mode."
>>
>> This worked well with TPP 4.5.2, but now it isn't working.  Is the new
>> version of Tandem using different defaults for mods?  I am not specifying
>> any in the tandem.params file.  I get the following error: (relevant part
>> of pipeline output below)
>> .
>> .
>> .
>>
>> running: "C:/Inetpub/tpp-bin/ASAPRatioPeptideParser 
>> "TEST-HL-interact.pep.xml" -lnK -b -F -C -Z -r0.5 -S"
>> K 170.142 176.18
>> n 43.0548 49.0928
>> error: cannot have variable modified n in static mode
>>
>> command "C:/Inetpub/tpp-bin/ASAPRatioPeptideParser 
>> "TEST-HL-interact.pep.xml" -lnK -b -F -C -Z -r0.5 -S" failed: Operation not 
>> permitted
>>
>> command "C:/Inetpub/tpp-bin/ASAPRatioPeptideParser 
>> "TEST-HL-interact.pep.xml" -lnK -b -F -C -Z -r0.5 -S" exited with non-zero 
>> exit code: 1
>> QUIT - the job is incomplete
>>
>> command "c:\Inetpub\tpp-bin\xinteract -NTEST-HL-interact.pep.xml -p0.05 -l6 
>> -Op -I4 -I5 -A-lnK-b-F-C-Z-r0.5-S 
>> 120827_S-Aorta_GuanidineExtract-RABA-QC01.tandem.pep.xml 
>> c:/Inetpub/wwwroot/ISB/data/projects/S_Aorta/120827_Guanidine/light/120827_S-Aorta_GuanidineExtract-RABA-QC01.tandem.pep.xml"
>>  failed: Operation not permitted
>>
>>
>>
>> Thanks in advance for any help.
>>
>> Brian
>>
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