I just wanted to add that I checked on previously analyzed samples, and 
that apparently, with the same settings and experimental conditions, I got 
the expected distribution of 2+ and 3+ peptides before. This sample was 
acquired on march of this year, probably with an earlier version of TPP. 
After that acquisition, I checked my results, and I have only identified 2+ 
OR 3+ peptides in the pep.shtml files (I didn't realize back then on this 
peculiar observation). Could someone check if this has happened in your set 
of results?

Thanks!

Alex

On Tuesday, 3 July 2012 17:03:05 UTC-3, Alex wrote:
>
> Hi TPP community!
>
> I have an interesting puzzle that I can not figure out!!! I'm running a 
> 2D-LC-MS/MS experiment of a yeast lysate (peptides labeled by 
> dimethylation) on a QqLIT (QTrap 3000) instrument acquiring on IDA mode.
> I'm analyzing the data on the TPP using X!Tandem search engine with the 
> params file attached, and interestingly, ALL the peptides identified on the 
> pep.shtml file are derived from triply charged precursors!!! (ALL of them). 
> When I check the prot.shtml file all the identified peptides have the 
> 3_SEQUENCE nomenclature (my understanding is that the 3 identifies the 
> charge on the precursor, right?). When I look at the raw data (.wiff) file 
> on Analyst software, I can easily estimate that most (probably a little 
> more than half) of the MS/MS was actually performed on CLEARLY doubly 
> charged precursors, according the the ER-MS (enhanced resolution MS 
> acquired on the LIT)... so it seem very weird that I dont find a single 
> peptide to spectrum match (PSM) from a doubly charged precursor. Just 
> wondering if I might have accidentally changed a parameter on the params 
> file (.xml).
>
> Has anybody crossed upon this observation before??? Anybody can help me???
>
> Thanks!!!
>
> Alex
>

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