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transproteomic-pipeline's Introduction

Update

This project and workflow has been merged into the WOMBAT pipelines.

Please go to https://github.com/wombat-p/WOMBAT-Pipelines

Workflow to analyze label-free data with the trans-proteomic pipeline

This workflow is based on a docker image which is available at the docker hub (https://hub.docker.com/repository/docker/wombatp/transproteomic-pipeline) and is downloaded automatically. If you want to build your own docker image, be aware that it needs to be named wombatp/transproteomic-pipeline:dev or change the name in the configuration file.

Content

Nextflow folder: Implementation of workflow

Results folder: Output from running the UPS data set

Getting started

Here is a little script that can be used to test the workflow execution. This script assumes that Docker is available on your system and is targeting Debian/Ubuntu linux distributions.

# Install Java
sudo apt-get install openjdk-8-jdk

# Fix docker permission issue (https://stackoverflow.com/questions/48957195/how-to-fix-docker-got-permission-denied-issue)
# Note: this allows to run docker as non-root user
sudo usermod -aG docker $USER
newgrp docker

# Fetch scripts and data
git clone https://github.com/wombat-p/Transproteomic-Pipeline.git
cd ./Transproteomics-Pipeline/Nextflow/data

# Nextflow
cd ..
curl -s https://get.nextflow.io | bash

# Worfklow
./nextflow run main.nf -profile docker,test

Run benchmarking data set

You should have successfully tested the workflow using the procedure above.

Download the raw files from PRIDE: http://proteomecentral.proteomexchange.org/cgi/GetDataset?ID=PXD001819

Run the workflow with the following command and parameters after changing RAWFOLDER to the folder where the raw files are located. You will also need to place the files yeast_UPS.fasta and pxd001819.txt into in the current folder. These files is given in the Results folder: https://github.com/veitveit/IS_Benchmarking/tree/master/Proline/Results

Also adjust the parameter values max_cpus and max_memory to the computing power you have available.

nextflow run main.nf --raws 'RAWFOLDER/*.raw' --fasta yeast_UPS.fasta --miscleavages 2 --fragment_mass_tolerance 0.8 \
--precursor_mass_tolerance 5 --enzyme 'Trypsin/P' --variable_mods 'Oxidation of M,Acetylation of protein N-term' \
--fdr_peptide_threshold 0.05 --quantification_fdr 0.01 --experiment_design pxd001819.txt --max_cpus 8 --max_memory \
8GB -profile docker -with-report -with-trace -with-timeline


transproteomic-pipeline's People

Contributors

veitveit avatar julianu avatar flevander avatar

Watchers

James Cloos avatar  avatar

Forkers

julianu

transproteomic-pipeline's Issues

Workflow fails at db_search_comet step

Step to reproduce :

./nextflow run main.nf -profile docker,test

dataset used
https://github.com/wombat-p/Transproteomic-Pipeline/tree/dev/Nextflow/data_test

error

Error executing process > 'db_search_comet (2)'

Caused by:
  Process `db_search_comet (2)` terminated with an error exit status (139)

Command executed:

  comet -P"comet.params" -N"OVEMB150205_12" -D"ABRF_iPRG_2012_target.fasta" "/home/gelonch/dev/jupyterlab_root/wombatp/Transproteomic-Pipeline/Nextflow/work/4e/a9bba0405ad47875141d6ef75845ad/OVEMB150205_12.mzML"

Command exit status:
  139

Command output:
   Comet version "2019.01 rev. 4"

Command error:
  .command.sh: line 2:     8 Segmentation fault      (core dumped) comet -P"comet.params" -N"OVEMB150205_12" -D"ABRF_iPRG_2012_target.fasta" "/home/gelonch/dev/jupyterlab_root/wombatp/Transproteomic-Pipeline/Nextflow/work/4e/a9bba0405ad47875141d6ef75845ad/OVEMB150205_12.mzML"

Work dir:
  /home/gelonch/dev/jupyterlab_root/wombatp/Transproteomic-Pipeline/Nextflow/work/dd/8a89851dadfabe44a8e5b37429ecc0

Tip: view the complete command output by changing to the process work dir and entering the command `cat .command.out`

PeptideProphet crashes

When running the workflow on an old laptop now running the latest Linux Mint, I run into a problem at the first PeptideProphet execution. Could it be I need more RAM? There is a warning "WARNING: Your kernel does not support swap limit capabilities or the cgroup is not mounted. Memory limited without swap.". The system has 8 GB.

Any idea what could be the issue? I could switch to a more modern system, but I wanted to try something different. And it seems it got pretty far, at least with Docker, Nextflow, msconvert and Comet...

Magnus

magnus@HP-EliteBook-850-G1:~/WOMBAT/Transproteomic-Pipeline/Nextflow$ ./nextflow run main.nf --raws 'data/PXD001819/*.raw' --fasta yeast_UPS.fasta --miscleavages 2 --fragment_mass_tolerance 0.8 \
> --precursor_mass_tolerance 5 --enzyme 'Trypsin/P' --variable_mods 'Oxidation of M,Acetylation of protein N-term' \
> --fdr_peptide_threshold 0.05 --quantification_fdr 0.01 --experiment_design pxd001819.txt --max_cpus 8 --max_memory \
> 8GB -profile docker -with-report -with-trace -with-timeline
N E X T F L O W  ~  version 21.04.1
Launching `main.nf` [admiring_lamarr] - revision: 55ccddcdf3
executor >  local (57)
[de/090b69] process > convert_raw_mzml (27)       [100%] 27 of 27 ✔
[a3/e81ac1] process > set_comet_configuration (1) [100%] 1 of 1 ✔
[e6/1e3b11] process > db_search_comet (27)        [100%] 27 of 27 ✔
[31/e54531] process > run_peptideprophet (2)      [  0%] 0 of 27
[-        ] process > run_proteinprophet          -
[-        ] process > run_stpeter                 -
[-        ] process > convert_stpeter             -
[-        ] process > run_merge_quant             -
Error executing process > 'run_peptideprophet (1)'

Caused by:
  Process `run_peptideprophet (1)` terminated with an error exit status (139)

Command executed:

  xinteract -N"UPS1_2500amol_R3.pep.interact.pep.xml" -p"0.05"  -l"8" -THREADS=4 -PPM -O -D"yeast_UPS.fasta" "/home/magnus/WOMBAT/Transproteomic-Pipeline/Nextflow/work/a0/6f71ea17ad8396abc0d55b2b705a1f/UPS1_2500amol_R3.pep.xml"

Command exit status:
  139

Command output:
  
  xinteract (TPP v5.2.0 Flammagenitus, Build 201904252338-7913 (Linux-x86_64))
   using THREADS=4 for iProphet...
   PPM mode in Accurate Mass Model ...
  
  running: "/usr/local/tpp/bin/InteractParser 'UPS1_2500amol_R3.pep.interact.pep.xml' '/home/magnus/WOMBAT/Transproteomic-Pipeline/Nextflow/work/a0/6f71ea17ad8396abc0d55b2b705a1f/UPS1_2500amol_R3.pep.xml' -D'yeast_UPS.fasta' -L'8'"
   file 1: /home/magnus/WOMBAT/Transproteomic-Pipeline/Nextflow/work/a0/6f71ea17ad8396abc0d55b2b705a1f/UPS1_2500amol_R3.pep.xml
   processed altogether 37673 results
  command completed in 1 sec 
  
  Use database specified by -D option: yeast_UPS.fasta. Skip DatabaseParser step.
  
  running: "/usr/local/tpp/bin/RefreshParser 'UPS1_2500amol_R3.pep.interact.pep.xml' 'yeast_UPS.fasta'"
  
    - Searching the tree...
    - Linking duplicate entries...
    - Printing results...
  
  command completed in 0 sec 
  
  running: "/usr/local/tpp/bin/PeptideProphetParser 'UPS1_2500amol_R3.pep.interact.pep.xml' MINPROB=0.05 PPM"
  using PPM mass difference
   (Comet)
  MS Instrument info: Manufacturer: UNKNOWN, Model: UNKNOWN, Ionization: UNKNOWN, Analyzer: UNKNOWN, Detector: UNKNOWN
  
  command completed in 0 sec 
  
  running: "/usr/local/tpp/bin/ProphetModels.pl -i UPS1_2500amol_R3.pep.interact.pep.xml"
  Analyzing UPS1_2500amol_R3.pep.interact.pep.xml ...
  Parsing search results "/home/magnus/WOMBAT/Transproteomic-Pipeline/Nextflow/work/63/2be92a1bc5038200aa08596169f5fd/UPS1_2500amol_R3 (Comet)"...
    => Found 0 hits. (0 decoys, 0 excluded)
    => Total so far: 0 hits. (0 decoys, 0 excluded)
  command completed in 0 sec 
  
  running: "/usr/local/tpp/cgi-bin/PepXMLViewer.cgi -I UPS1_2500amol_R3.pep.interact.pep.xml"
  
  command "/usr/local/tpp/cgi-bin/PepXMLViewer.cgi -I UPS1_2500amol_R3.pep.interact.pep.xml" exited with non-zero exit code: 35584
executor >  local (57)
[de/090b69] process > convert_raw_mzml (27)       [100%] 27 of 27 ✔
[a3/e81ac1] process > set_comet_configuration (1) [100%] 1 of 1 ✔
[e6/1e3b11] process > db_search_comet (27)        [100%] 27 of 27 ✔
[e6/7ddc27] process > run_peptideprophet (1)      [  4%] 1 of 27, failed: 1
[-        ] process > run_proteinprophet          -
[-        ] process > run_stpeter                 -
[-        ] process > convert_stpeter             -
[-        ] process > run_merge_quant             -
Error executing process > 'run_peptideprophet (1)'

Caused by:
  Process `run_peptideprophet (1)` terminated with an error exit status (139)

Command executed:

  xinteract -N"UPS1_2500amol_R3.pep.interact.pep.xml" -p"0.05"  -l"8" -THREADS=4 -PPM -O -D"yeast_UPS.fasta" "/home/magnus/WOMBAT/Transproteomic-Pipeline/Nextflow/work/a0/6f71ea17ad8396abc0d55b2b705a1f/UPS1_2500amol_R3.pep.xml"

Command exit status:
  139

Command output:
  
  xinteract (TPP v5.2.0 Flammagenitus, Build 201904252338-7913 (Linux-x86_64))
   using THREADS=4 for iProphet...
   PPM mode in Accurate Mass Model ...
  
  running: "/usr/local/tpp/bin/InteractParser 'UPS1_2500amol_R3.pep.interact.pep.xml' '/home/magnus/WOMBAT/Transproteomic-Pipeline/Nextflow/work/a0/6f71ea17ad8396abc0d55b2b705a1f/UPS1_2500amol_R3.pep.xml' -D'yeast_UPS.fasta' -L'8'"
   file 1: /home/magnus/WOMBAT/Transproteomic-Pipeline/Nextflow/work/a0/6f71ea17ad8396abc0d55b2b705a1f/UPS1_2500amol_R3.pep.xml
   processed altogether 37673 results
  command completed in 1 sec 
  
  Use database specified by -D option: yeast_UPS.fasta. Skip DatabaseParser step.
  
  running: "/usr/local/tpp/bin/RefreshParser 'UPS1_2500amol_R3.pep.interact.pep.xml' 'yeast_UPS.fasta'"
  
    - Searching the tree...
    - Linking duplicate entries...
    - Printing results...
  
  command completed in 0 sec 
  
  running: "/usr/local/tpp/bin/PeptideProphetParser 'UPS1_2500amol_R3.pep.interact.pep.xml' MINPROB=0.05 PPM"
  using PPM mass difference
   (Comet)
  MS Instrument info: Manufacturer: UNKNOWN, Model: UNKNOWN, Ionization: UNKNOWN, Analyzer: UNKNOWN, Detector: UNKNOWN
  
  command completed in 0 sec 
  
  running: "/usr/local/tpp/bin/ProphetModels.pl -i UPS1_2500amol_R3.pep.interact.pep.xml"
  Analyzing UPS1_2500amol_R3.pep.interact.pep.xml ...
  Parsing search results "/home/magnus/WOMBAT/Transproteomic-Pipeline/Nextflow/work/63/2be92a1bc5038200aa08596169f5fd/UPS1_2500amol_R3 (Comet)"...
    => Found 0 hits. (0 decoys, 0 excluded)
    => Total so far: 0 hits. (0 decoys, 0 excluded)
  command completed in 0 sec 
  
  running: "/usr/local/tpp/cgi-bin/PepXMLViewer.cgi -I UPS1_2500amol_R3.pep.interact.pep.xml"
  
  command "/usr/local/tpp/cgi-bin/PepXMLViewer.cgi -I UPS1_2500amol_R3.pep.interact.pep.xml" exited with non-zero exit code: 35584
  QUIT - the job is incomplete

Command error:
  WARNING: Your kernel does not support swap limit capabilities or the cgroup is not mounted. Memory limited without swap.
  WARNING: Unable to open file: /home/magnus/WOMBAT/Transproteomic-Pipeline/Nextflow/work/63/2be92a1bc5038200aa08596169f5fd/UPS1_2500amol_R3.mzML cannot correct scan numbers!
  WARNING: Unable to open file: /home/magnus/WOMBAT/Transproteomic-Pipeline/Nextflow/work/63/2be92a1bc5038200aa08596169f5fd/UPS1_2500amol_R3.mzML cannot correct scan numbers!
  INFO: Results written to file: UPS1_2500amol_R3.pep.interact.pep.xml
    - Building Commentz-Walter keyword tree...Segmentation fault (core dumped)

Work dir:
  /home/magnus/WOMBAT/Transproteomic-Pipeline/Nextflow/work/e6/7ddc27fbb0efcfac3813f8d87eb210

Tip: you can try to figure out what's wrong by changing to the process work dir and showing the script file named `.command.sh`


Oops .. something went wrong
WARN: Killing pending tasks (1)

magnus@HP-EliteBook-850-G1:~/WOMBAT/Transproteomic-Pipeline/Nextflow$

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