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)

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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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