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No decoys found in the FASTA file? #1675

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rolivella opened this issue Jul 16, 2024 · 7 comments
Closed

No decoys found in the FASTA file? #1675

rolivella opened this issue Jul 16, 2024 · 7 comments

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@rolivella
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Hello

I formatted a Human FASTA file using my own scripts for creating the decoy and Philosopher but I get from FragPipe this error:

2024-07-16 11:01:27,686 ERROR - No decoys found in the FASTA file.

I reviewed the FASTA and it seems OK and following the guidelines seen at the Philospher doc. Do you know why this error?

Thanks!

The file: sp_human_decoy_cont_formatted.fasta.zip

@fcyu
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fcyu commented Jul 16, 2024

Could you share your workflow file!

Thanks,

Fengchao

@rolivella
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Yes, here it is: fragpipe-220.workflow.zip

@fcyu
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fcyu commented Jul 16, 2024

You should change the decoy tag to database.decoy-tag=DECOY_:

database.decoy-tag=rev_

Best,

Fengchao

@rolivella
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@fcyu I reopen the issue because after running the fragpipe I got some unexpected messages and stopped before ending properly. I think that the problem is that it cannot run PeptideProphet:

time="15:54:08" level=info msg="Executing PeptideProphet  v5.1.1"
Unknown file type. No file loaded./home/proteomics/mygit/atlas-runs/520c5201-cc4e-4e32-a566-8504c41c27cc/59/571dd10648593fae17bab5703e7f3c/output/1_1/2024MQ024_MSGA_052_01_25pto.raw
Unknown file type. No file loaded./home/proteomics/mygit/atlas-runs/520c5201-cc4e-4e32-a566-8504c41c27cc/59/571dd10648593fae17bab5703e7f3c/output/1_1/2024MQ024_MSGA_052_01_25pto.raw

I searched for a similar error in the issues and I found this one:

#1556

That was related with precisely the decoy sequences but I think that in my case they are OK...Any idea? This is my log:

fragpipe22_manual_decoy.log.zip

Thanks!

@rolivella rolivella reopened this Jul 16, 2024
@fcyu
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fcyu commented Jul 16, 2024

The warnings you found won't affect the results. The crash was because the PeptideProphet models failed, and then there was no IDs. If you are doing a closed search, why not using Percolator?

Iterations: .........10.........20....WARNING: Mixture model quality test failed for charge (1 ).
.
WARNING: Mixture model quality test failed for charge (2 ).
WARNING: Mixture model quality test failed for charge (3 ).
WARNING: Mixture model quality test failed for charge (4 ).
WARNING: Mixture model quality test failed for charge (5 ).
WARNING: Mixture model quality test failed for charge (6 ).
WARNING: Mixture model quality test failed for charge (7 ).

Best,

Fengchao

@rolivella
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Ok, I see. Do you mean percolator.run-percolator=true? I did it and got the same error log: fragpipe22_manual_decoy_v2.log.zip

@fcyu
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fcyu commented Jul 16, 2024

You still have PeptideProphet running

peptide-prophet.run-peptide-prophet=true
percolator.run-percolator=false

Best,

Fengchao

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