Frederik,

Many thanks for this information, I hadn't come across the Proteios 
convertor. Unfortunately we're working now with SILAC data where we need 
the MS1 for quantitation, but the workflow might be useful for other 
data. I have previously extracted EMRT information from PLGS XML files 
in MS^e experiments, but hadn't thought to look at the XML files for DDA.

Thanks,

DT

fredrik wrote:
> Hi Dave,
>
> An option could be to run PLGS and then convert the PLGS XML files to
> mzML using the Proteios SE converter. Using this workflow the spectrum
> ids and retention times are retained. However, the level 1 MS data is
> lost, so it might not be ideal for your workflow, apart from the
> identifications. But you'll probably gain lock mass correction and
> spectrum summing, compared to the massWolf workflow. The standalone
> converter is found here:
> http://dev.thep.lu.se/fp6-prodac/browser/trunk/mzML/mzMLconverters.zip
> The input PLGS XML files are found in subdirectories of the PLGS
> project directory, and are all named MassSpectrum.xml.
> Here is a perl script that renames the PLGS files to the name of the
> raw data files:
> http://www.proteios.org/trac/wiki/GelBasedReport
>
> Regards
>
> Fredrik
>
> On Sep 14, 1:37 pm, dctrud <[email protected]> wrote:
>   
>> Dear all, another query r.e. Waters QTOF data....
>>
>> We have a QTOF Premier which is now being used for SILAC experiments,
>> and we want to quantify using ASAPRatio / Xpress. Have no problems
>> doing this in our pipeline using Orbitrap data, but the QTOF data is
>> causing headaches. Up until now we've mostly been using MassWolf just
>> to get MS data into .mzXML for purposes other than ID & quant, PLGS
>> having been the preferred analysis tool for the instrument.
>>
>> If we use .mzXML files and feed them to Mascot/OMSSA/Tandem + TPP we
>> cannot get anywhere near the number of IDs as with .pkl files
>> generated using PLGS. We've tried conversion to mzXML using MassWolf
>> and msconvert, applying lockmass correction in MassLynx prior to
>> conversion, and comparing acquisition of MS/MS scans in centroid mode,
>> vs acquisition in profile mode converted to centroid using MassLynx.
>> An example of the differences in IDs:
>>
>> RAW->PLGS->PKL->Mascot->TPP: 874 peptide IDs (1% FDR)
>> RAW->MassWolf->mzXML->mgf->Mascot->TPP: 279 peptide IDs (1 % FDR)
>>
>> No matter what combination of converter / pre-processing in MassLynx
>> we try, the number of IDs is never above 1/3 of that we get from a PKL
>> file. We've also tried changes to the acquisition methods, without
>> success.
>>
>> Is anyone able to provide a MassLynx method file, and/or procedure for
>> pre-processing / .mzXML conversion they are using successfully with a
>> QTOF Premier or similar instrument?
>>
>> Any help would be gratefully appreciated. Many Thanks,
>>
>> DT
>>     
>
> >
>   


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