Hi David

Apologies for the hassle! I have uploaded the .d files in an unzipped 
format under TPP_diagnosis/TPP_diagnosis. Let me know if this works instead?

Best
Shagun

On Tuesday, August 13, 2024 at 3:09:37 PM UTC-4 David Shteynberg wrote:

> Hello Shagun,
>
> I downloaded the larger zip file twice and tried to decompress, but each 
> time it told me there was a corruption in the zip file.  The smaller files 
> downloaded fine.  Can you check your file and upload again?
>
> Thanks!
> -David
>
> On Tue, Aug 13, 2024 at 9:47 AM Shagun Gupta <[email protected]> wrote:
>
>> Hi David
>>
>> I have attached a link to the following files 
>> - one replicate per condition .d file (file starting with "C_" had IDs 
>> but no values that could be extracted for quantification)
>> - comet parameter file I used
>> - fasta file I ran the search with.
>>
>> Let me know if I can share anything else!
>>
>> Box_link_diagnosis 
>> <https://cornell.box.com/s/5y2uak2twszajaniaqpnevxn4zdneu54>
>> Best
>> Shagun
>>
>> On Tuesday, August 13, 2024 at 11:05:42 AM UTC-4 David Shteynberg wrote:
>>
>>> Hello Shagun,
>>>
>>> Thank you for the detailed report.  If you are able, please first 
>>> compress (into a zip or similar) and then share some of the problem .d 
>>> files so I can try to replicate this issue on my computer before I offer 
>>> any suggestions.  
>>>
>>> Cheers!
>>> -David
>>>
>>> On Tue, Aug 13, 2024 at 7:50 AM Shagun Gupta <[email protected]> wrote:
>>>
>>>> Hi all
>>>>
>>>> I have been experiencing issues processing .d files obtained from a 
>>>> Bruker timsTOF HT in DDA-LFQ mode, more specifically getting 
>>>> quantification 
>>>> - precursor intensity - per spectra. I am using TPP V6.3.3 Arcus on a 
>>>> windows computer. 
>>>>
>>>> Details
>>>> - There are 12 .d files (4 repeats of 3 conditions) composed of a human 
>>>> with yeast proteome spike in at different ratios.
>>>> - Converted to .mzXML using msconvert
>>>> - Searched with COMET and a search database taken from UniProt for Homo 
>>>> sapiens+Yeast
>>>> - Processed with PeptideProphet (filtered at probability associated 
>>>> with 1% FDR), XPRESS, ProteinProphet. Ran with -PREC flag 
>>>> (PeptideProphet), 
>>>> -i flag (XPRESS)
>>>> - Want to do hypothesis testing (comparing the three conditions 
>>>> pairwise) using MSstats. So require raw precursor intensity values to make 
>>>> a file that can be used as input to MSstats.
>>>>
>>>> Unfortunately after trying the above, and a few more things, while I 
>>>> get a large number of PSMs passing FDR (~30k), a large proportion of them 
>>>> do not have any precursor intensity value (<1k have some "light area" 
>>>> values). Using the "light area" values also does not give expected results 
>>>> (its a benchmarking dataset and processing with MSFragger gave excellent 
>>>> results that align with expected ratios etc.). Could you suggest things I 
>>>> could be doing differently to get the right results? (I imagine it might 
>>>> have something to do with the initial conversion to mzXML itself?) 
>>>>
>>>> Happy to share any other details needed!
>>>>
>>>> Best
>>>> Shagun
>>>>
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