Hi Ankit, both strategies would likely work okay. I would prefer first
combining all your data with iProphet and then creating the library with
SpectraST. An issue to consider: if you combine several libraries that have
been processed or thresholded individually, the final FDR will be higher
for the combined dataset than the individual ones. This is because the
correct identifications will mostly be in common, but the false
identifications will be mostly different in the different experiments and
so they will accumulate. So combining everything first with iProphet will
give you a better understanding of your FDR.



Regards,

Eric





*From:* [email protected] <[email protected]>
*On Behalf Of *Ankit Balhara
*Sent:* Thursday, March 14, 2019 11:47 PM
*To:* spctools-discuss <[email protected]>
*Subject:* [spctools-discuss] Combining the data on two tissues in one
library



Hi,



I have acquired the DDA and DIA data on various tissues, and trying to
build a spectral library by combining the DDA results of all tissues, that
can be used further for the DIA data analysis. I am trying two approaches-
one assay library generation with SpectraST and other using skyline to
build the library. Now, I confused, how should I proceed.



Should I combine the peptideprophet results of various tissues by using
iProphet?

Or, create assay libraries for each tissue separately using the SpectraST
and then import them separately in a common Skyline document?



I would be very grateful for your help.



Thanks in advance.

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