Hi Ludovic,

It seems the file you uploaded lgillet_pepxml_for_TPP4.3.rar is
corrupted.  At least I am unable to open it. Please upload again.

Thanks,
-David

On Wed, May 19, 2010 at 2:54 AM, lgillet <[email protected]> wrote:
> Hi David, Hi Natalie,
>
> I just posted the 4 pepxml files which give me the most striking
> differences in results between TPP-V4.0 and TPP-V4.3:
> lgillet_pepxml_for_TPP4.3.rar. I also posted the results
> (interact.pep.xml) which I obtain from running TPP-V4.0, TPP-V4.3 and
> TPP-V4.3 on scrambled file order (file #4>#3>#2>#1): lgillet_interact-
> results.rar.
> I really tried my best to figure out what the problem could be.
> Maybe you could re-run the same analyses (TPP-V4.0, TPP-V4.3, TPP-V4.3
> with the scrambled file order) and let me know if you confirm my
> results or if there is something wrong maybe with the compiled version
> we have on our server (could still be a possibility).
> Finally, to answer Natalie's question, the differences are quite
> dramatic (to my opinion) between V4.0 and V4.3 (I would not have
> worried about 1-2% differences in IDs), but here, I am passing from 1%
> decoy (V4.0) to 23% decoy (V4.3) hits (at the same proba > 0.9). Also
> the number of unique peptides reported by V4.0 and V4.3 is quite
> different (2150 and 3161 resp.). Finally, many decoy hits pulled up in
> V4.3 with a prob>0.9 have actually a very bad MS/MS spectrum and a
> very low prob<0.01 (only reported if you use -p0 option) on V4.0.
>
> Have a look at those MS/MS spectra for example:
>
> 20100422_04_control_07.c.07700.07700.4
> 20100422_04_control_07.c.02864.02864.3
>
> Let me know if you need any extra information.
>
> Thanks a lot for your help on that.
>
> Best,
>
> Ludovic
>
> On May 18, 11:21 pm, Natalie Tasman <[email protected]>
> wrote:
>> Ludovic,
>>
>> Go ahead and post the files to the newsgroup's file area
>> (http://groups.google.com/group/spctools-discuss/files), and hopefully
>> one of the validation experts will take a look.
>>
>> I will point out that PeptideProphet uses random initialization for
>> it's curve fitting (EM algorithm).  So it's not out of the question
>> that you'd see some small differences between runs on the same data
>> files, regardless of the order.  Can you provide some measure of the
>> differences between runs for the reordered datasets?
>>
>> -Natalie
>>
>>
>>
>> On Tue, May 18, 2010 at 4:35 AM, lgillet <[email protected]> wrote:
>> > Hi everybody,
>> > I recently encountered a "bug" I think when people in my lab installed
>> > the newest TPP (v4.3 JETSTREAM rev 1, Build 201004201202 (linux)),
>> > especially when I try to confront the result to v4.0 which was our
>> > former "benchmark" version.
>> > When searching the same 4 pep.xml files with v4.0 and v4.3, I get an
>> > incredible difference in decoy hits number. For example, with v4.0,
>> > p>0.9, I would get my "regular" 1% decoy, while with v4.3, p>0.9, I
>> > get above 25% of decoys?!??
>> > All the interact are run with the following options: xinteract -OApld -
>> > ddecoy *.pep.xml
>> > I could nail down the "problem" to the PeptideProphetParser which
>> > behaves very differently between v4.0 and v4.3, while InteractParser
>> > (which introduces the "is_rejected=1" tags) and RefreshParser do not
>> > influence the results.
>> > But at the moment, I do not know if it is an issue of the decoy
>> > statistical distribution of prophet or not...
>>
>> > One more thing that makes me even more suspicious is the fact that,
>> > only with TPP version 4.3, if you search those files in a difference
>> > order (let say: xinteract file1 file2 file3 Vs xinteract file3 file2
>> > file1), you do get differences in the results as well?!?
>>
>> > I am willing to send the 4 pepxml where those observations are the
>> > most critical to David or Luis or anybody interested, but I truly
>> > believe that there might be something going wrong with the TPP v4.3.
>>
>> > Let me know to whom I should post the files.
>>
>> > Best regards,
>>
>> > Ludovic
>>
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