Thank you so much for the quick reply! I did not realize Comet did this. I 
will change my params file and run the search again.

Thanks again

On Thursday, March 1, 2018 at 3:55:35 PM UTC-5, Jimmy Eng wrote:
>
> Francesca,
>
>    Quick answer:  set "decoy_search = 0" because your FASTA already has 
> decoy entries in it.
>
> The "decoy_search" parameter controls whether or not you have Comet 
> generate decoys "on-the-fly".  Since your database already has decoy 
> entries, you need to set "decoy_search = 0".  Otherwise, you're telling 
> Comet to take all of your sequences in your FASTA, including your decoy 
> entries, and create a new internally-generated decoy peptide for each 
> peptide it reads from the database.
>
> So if you have a decoy protein "DECOY9389", you're having Comet take those 
> decoy peptides and score them.  But by setting "decoy_entry = 1", you're 
> also directing Comet to generate new decoys of those decoy peptides.  When 
> Comet generates new decoys, it appends "DECOY_", based on the 
> "decoy_prefix" parameter, to the beginning of the existing protein name.  
> In this example, it would be "DECOY_DECOY9389".
>
>
> Jimmy
>
> On Thu, Mar 1, 2018 at 12:41 PM, Francesca Barry <[email protected] 
> <javascript:>> wrote:
>
>> Hello,
>>
>> I just realized a possible issue I've been having when using Comet to 
>> perform a concatenated decoy search on my .mzml file. I just noticed that 
>> the decoy matches are being displayed/outputted in different ways.
>>
>> In both the .pep.xml and .pin output files (I need to run Percolator 
>> after), the decoy hits are being displayed as one of the 3 forms (i.e. all 
>> 3 forms are present in the same file):
>>
>> DECOYsp|Q8IZQ1|WDFY3_HUMAN
>> DECOY6157
>> DECOYDECOY9389
>>
>> When I check back in the .fasta file, the DECOYDECOY results are 
>> reversing the decoys (so they become the original forward peptides), which 
>> is wrong. I'm not sure why I'm getting all 3 forms, especially the 
>> DECOYDECOY matches in the output files. My .fasta is a concatenated version 
>> of the forward and reverse (decoy) peptides.
>>
>> What's more interesting is that when I convert the .pep.xml file to .mzID 
>> so I can further process the results, the .mzID does not contain any 
>> DECOYDECOY entries.
>>
>> Very confused as to what is happening and any help would be much 
>> appreciated. I would especially like to sort out this issue I'm having with 
>> Comet.
>>
>> Thanks,
>>
>> Francesca
>>
>> PS. I have also uploaded the .params file for Comet in case there is an 
>> error in there.
>>
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