It should be number(decoys)/number(targets) irrespective of
concatenated or separate searches.

On Wed, Nov 24, 2010 at 5:20 AM, Amit Yadav <[email protected]> wrote:
> Hi,
>
> From what I understand, the simplest ways to calculate FDR using a
> Target-Decoy search are:-
>
> 1. Run a Concatenate target and decoy database search. The formula
> would be :    2  *number(decoys)/ number(target+decoys) - See Elias
> and Gygi , Nat Methods. 2007 Mar;4(3):207-14.
>
> 2. Run Separate target and decoy searches. The formula would be:
> number(decoys)/number(targets). See Kall et al J Proteome Res. 2008
> Jan;7(1):29-34
>
> I think your formula should be the first one. Members, please correct
> me if I am wrong.
>
> Regards,
>
> Amit Kumar Yadav
> Senior Research Fellow (SRF-CSIR)
> IGIB, New Delhi (India)
>
> MassWiz Web server
> MassWiz sourceforge project
> MassWiki
>
>
>
> On Wed, Nov 24, 2010 at 6:39 PM, Bjorn <[email protected]> wrote:
>>
>> Hello all,
>>
>> I am trying to find my way through the different interpretations and
>> calculations of the FDR, but I'm a bit lost where it comes to the
>> PeptideProphet.
>>
>> So far, I know that the simplest calculation is (decoy_hits/
>> target_hits). So, I ran X!tandem on my dataset (target DB +
>> concatenated decoy DB) and then ran PeptideProphet on the resulting
>> pep.xml file (not setting the decoy option!).
>> I then set a certain probability threshold (say 0.85) and looked at
>> the number of spectra (e.g. 1200 of 1500). Setting the protein text to
>> decoy gives me, e.g. 50 hits. The FDR for a prob. of 0.85 would then
>> be 50/1150. If I look at the table displayed in the PeptideProphet,
>> the FDR I found is almost spot-on with the number in the table. The
>> thing is that if I lower my probabilty threshold, the FDRs in the
>> table stay relatively low, while the FDRs I calculate with the above
>> formula go up. Is there any extra correction being done in the
>> PeptideProphet (like correct for the percentage of incorrect target
>> hits (PIT) as described by Kall et al in 2008)?
>> Or am I missing something vital here?
>>
>> Furthermore, how is the estimated number of correct assignments
>> calculated? I can't seem to find the correct formula to come to that
>> number? I assume it is related to the surface under the curve plotted
>> with the correct identifications?
>>
>> Many questions, but I hope someone can help me with this.
>>
>> Best regards,
>>
>> bjorn
>>
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