Thanks very much for your answers - that's very helpful!
Alastair

Am Freitag, 13. Dezember 2019 21:56:04 UTC+1 schrieb Eric Deutsch:
>
> Thanks for the clarification, Jimmy!
>
>  
>
> *From:* [email protected] <javascript:> <
> [email protected] <javascript:>> *On Behalf Of *Jimmy Eng
> *Sent:* Friday, December 13, 2019 8:27 AM
> *To:* [email protected] <javascript:>
> *Subject:* Re: [spctools-discuss] Interpretation of stats from pepXML 
> viewer
>
>  
>
> Regarding inputs and output counts to Comet, as Eric mentioned, there are 
> search parameters which affect whether or not a spectrum will be searched 
> at all.  These are: activation_method, digest_mass_range, 
> max_precursor_charge, minimum_peaks, minimum_intensity, and 
> remove_precursor_peak (which can affect minimum_peaks).  A query spectrum 
> that happens to be out of bounds for any of these parameters will not be 
> analyzed (is effectively ignored) and will not show up in the output.  Any 
> spectra that passes these filters will have an output entry in Comet's 
> pepXML output file, even if there's no matching peptide output.  Such blank 
> outputs can happen in cases where the database or tolerances are so small 
> that no peptides are scored or the spectrum is so poor that the xcorr score 
> is less than or equal to zero.  This is why you see 1752 spectra in the 
> pepXML after Comet searches of both your databases as that's the number of 
> spectra that actually get analyzed and is a function of the input and not 
> the database being searched.
>
>  
>
> On Fri, Dec 13, 2019 at 2:29 AM 'Alastair Skeffington' via 
> spctools-discuss <[email protected] <javascript:>> wrote:
>
> Hi,
>
>  
>
> Can someone explain what's happen here please:
>
>  
>
> Number of spectra in the .mgf file = 1856
>
> Number of spectra after conversion to mzXML = 1856
>
> Number of spectra in pepXML file after comet search: 1752
>
>  
>
> I then search against database A (DB-A) and database B (DB-B) and count 
> the number of spectra in the pepXML file as counts of "<spectrum_query 
> spectrum=". Yields 1752 in both cases.
>
>  
>
> I then run "xinteract -Neg.pepXML -PPM -dDECOY_ -OARP eg_pepp.pepXML" on 
> each of the files. Now when I search for the string "<spectrum_query 
> spectrum=" I get 481 hits for DB-A and 632 hits for DB-B. The pepXML view 
> then calculates the "Efficiency ID'd/searched" - so the denominator is 
> different for each database.
>
>  
>
> Can someone help to explain this behavior?
>
>  
>
>    - Why does the xinteract output present different numbers of records 
>    ("<spectrum_query spectrum= ...") for the two different databases, even 
>    though the number of records in the pepXML input was the same?
>    - Isn't the definition of 'Spectra searched' used by Petunia 
>    misleading? It's not the number of input spectra to the search engine.
>    - Presumably the loss of spectra between the input to comet and the 
>    output is due to loosing those spectra with absolutely no match to the 
>    database or decoys due to some internal threshold on the score for PSMs.
>
>  
>
> Any help would be hugely appreciated!
>
>  
>
> Thanks,
>
> Alastair
>
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