Hi Jimmy,

I was following Elias & Gygi's formula for concatenated searches. I was
unaware that the same formula is to be used for both searches. Please
provide me with the reference.

Regards,

*Amit Kumar Yadav *
Senior Research Fellow (SRF-CSIR)
IGIB, New Delhi (India)

*MassWiz Web server* <http://masswiz.igib.res.in>
* <http://masswiz.igib.res.in>**MassWiz sourceforge
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* 
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On Fri, Nov 26, 2010 at 12:03 AM, Jimmy Eng <[email protected]> wrote:

> 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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