It sounds like a problem with the way your decoys might be getting
generated.  How do you generate the decoy?

On Tue, Apr 4, 2017 at 8:59 AM, Chih-Chiang Tsou <[email protected]>
wrote:

> Hi David,
>
> Thanks for the prompt response.
> So what I am trying to do is to avoid decoy hit Blocking the possible true
> hit.
>
> The way I perform the analysis is to search data against a FASTA file
> which has reversed sequences appended. Then during PeptideProphet analysis
> I specify the decoy prefix and in the end use the probabilities for
> target/decoy hits to estimate FDR. One potential problem for me is that if
> a spectrum happens to match a decoy as best hit, with a very high score
> let's say XCorr 2.0, and the second hit with Xcorr 1.9, which si actually
> the true hit, would be discarded after PeptideProphet analysis.
>
> If PeptideProphet takes only top hit, could I do two separate searches
> using target-FASTA and Decoy-FASTA and specify two input files in
> PeptideProphet? Any suggestions?
>
>
> Thanks,
> Chih-Chiang
>
> On Tuesday, April 4, 2017 at 9:45:11 AM UTC-5, David Shteynberg wrote:
>>
>> No.  It uses the top hit only.
>>
>> On Tue, Apr 4, 2017 at 7:34 AM, Chih-Chiang Tsou <[email protected]>
>> wrote:
>>
>>> Hi
>>>
>>> Is there any option in xinteract to tell PeptideProphet take topN hits
>>> for each spectrum for probability calculation?
>>>
>>> Thanks,
>>> Chih-Chiang
>>>
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