diff --git a/refs/UP000005640_9606.fasta.gz b/refs/UP000005640_9606.fasta.gz new file mode 100644 index 0000000..cc4b02e Binary files /dev/null and b/refs/UP000005640_9606.fasta.gz differ diff --git a/run_fragpipe.sh b/run_fragpipe.sh deleted file mode 100644 index 5bd148f..0000000 --- a/run_fragpipe.sh +++ /dev/null @@ -1,87 +0,0 @@ -#!/bin/bash - -set -e -set -o pipefail - -# Set tools and input directories -input_dir=input -tools_dir=/fragpipe_bin/fragPipe-23.1/fragpipe/tools - -# Error out if no custom.fasta is provided -if [ ! -e "$input_dir/custom.fasta" ]; then - echo "Error: must supply a custom.fasta file in the input directory ($input_dir)" - exit 1 -fi - -echo "Adding decoys and contaminants to FASTA files.." -cd $input_dir - -# Initialize the Philosopher workspace -$tools_dir/Philosopher/philosopher-v5.1.1 workspace --init - -# Add decoys and contaminants to the custom FASTA file -gunzip -c UP000005640_9606.fasta.gz | $tools_dir/Philosopher/philosopher-v5.1.1 database --custom custom.fasta --add /dev/stdin --contam -mv *decoys-contam-custom.fasta.fas decoys-contam-custom-canonical.fasta - -# Clean intermediate files -$tools_dir/Philosopher/philosopher-v5.1.1 workspace --clean - -# Return to the previous directory -cd - - -# set cavatica directory -cavatica_dir=(/home/rstudio/fragpipe/data/projects/harenzaj/proteomics) - -if [ ! -d $cavatica_dir ]; then - echo "Error: path to cavatica project directory does not exist. Confirm that proteomics cavatica project is mounted to data/ directory" - exit 1 -fi - -# check if tmp directory exists, and create it if not -tmp_dir=(/home/rstudio/fragpipe/tmp) - -if [ ! -d $tmp_dir ]; then - mkdir -p $tmp_dir - echo "Temporary directory does not exist, creating..." -fi - -# clear contents of tmp directory, if not already empty -rm -rf $tmp_dir/* - -# copy mzML files to temporary directory, maintaining directory structure -echo "copying mzML files to temporary directory..." - -if ! [[ " $* " == *" --run_subset "* ]]; then - -cp -R $cavatica_dir/*CBTTC_PBT_Proteome* /home/rstudio/fragpipe/tmp/ - -else - -cp -R $cavatica_dir/01CBTTC_PBT_Proteome* /home/rstudio/fragpipe/tmp/ - -fi - -# unzip mzML files, force overwrite if files exist -echo "unzipping mzML files..." -gunzip -f /home/rstudio/fragpipe/tmp/*CBTTC_PBT_Proteome*/*mzML.gz - -# set workflow, manifest, results and tools directories -echo "Starting Fragpipe run..." - -wf=/home/rstudio/fragpipe/input/PDC000180customworkflow.workflow -manifest=/home/rstudio/fragpipe/input/PDC000180filesmanifest.fp-manifest - -if [[ " $* " == *" --run_subset "* ]]; then - -grep "01CBTTC_" $manifest > /home/rstudio/fragpipe/input/PDC000180filesmanifest.fp-manifest_sub -manifest=/home/rstudio/fragpipe/input/PDC000180filesmanifest.fp-manifest_sub - -fi - -res_dir=/home/rstudio/fragpipe/results -tools_dir=/fragpipe_bin/fragPipe-23.1/fragpipe/tools - -/fragpipe_bin/fragPipe-23.1/fragpipe/bin/fragpipe --headless --workflow $wf --manifest $manifest --workdir $res_dir --config-tools-folder $tools_dir - -# clear tmp directory -rm -rf $tmp_dir/* diff --git a/tumor-enriched-splicing/input/PDC000180customworkflow.workflow b/tumor-enriched-splicing/input/PDC000180customworkflow.workflow new file mode 100644 index 0000000..15329cf --- /dev/null +++ b/tumor-enriched-splicing/input/PDC000180customworkflow.workflow @@ -0,0 +1,327 @@ +# Workflow: PDC000180customworkflow + + +# Please edit the following path to point to the correct location. +# In Windows, please replace single '\' with '\\' +database.db-path=/home/rstudio/fragpipe/tumor-enriched-splicing/input/decoys-contam-custom-canonical.fasta + +crystalc.run-crystalc=false +database.decoy-tag=rev_ +diann.fragpipe.cmd-opts= +diann.generate-msstats=true +diann.heavy= +diann.library= +diann.light= +diann.medium= +diann.q-value=0.01 +diann.quantification-strategy=3 +diann.quantification-strategy-2=QuantUMS (high accuracy) +diann.run-dia-nn=false +diann.run-dia-plex=false +diann.run-specific-protein-q-value=false +diann.unrelated-runs=false +diann.use-predicted-spectra=true +diaumpire.AdjustFragIntensity=true +diaumpire.BoostComplementaryIon=false +diaumpire.CorrThreshold=0 +diaumpire.DeltaApex=0.2 +diaumpire.ExportPrecursorPeak=false +diaumpire.Q1=true +diaumpire.Q2=true +diaumpire.Q3=true +diaumpire.RFmax=500 +diaumpire.RPmax=25 +diaumpire.RTOverlap=0.3 +diaumpire.SE.EstimateBG=false +diaumpire.SE.IsoPattern=0.3 +diaumpire.SE.MS1PPM=10 +diaumpire.SE.MS2PPM=20 +diaumpire.SE.MS2SN=1.1 +diaumpire.SE.MassDefectFilter=true +diaumpire.SE.MassDefectOffset=0.1 +diaumpire.SE.NoMissedScan=1 +diaumpire.SE.SN=1.1 +diaumpire.run-diaumpire=false +fpop.fpop-tmt=false +fpop.label_control= +fpop.label_fpop= +fpop.region_size=1 +fpop.run-fpop=false +fpop.subtract-control=false +freequant.mz-tol=10 +freequant.rt-tol=0.4 +freequant.run-freequant=false +ionquant.excludemods= +ionquant.heavy= +ionquant.imtol=0.05 +ionquant.ionfdr=0.05 +ionquant.light= +ionquant.locprob=0.75 +ionquant.maxlfq=1 +ionquant.mbr=0 +ionquant.mbrimtol=0.05 +ionquant.mbrmincorr=0 +ionquant.mbrrttol=1 +ionquant.mbrtoprun=10 +ionquant.medium= +ionquant.minfreq=0 +ionquant.minions=2 +ionquant.minisotopes=2 +ionquant.minscans=3 +ionquant.mztol=10 +ionquant.normalization=1 +ionquant.peptidefdr=5 +ionquant.proteinfdr=5 +ionquant.requantify=1 +ionquant.rttol=0.4 +ionquant.run-ionquant=true +ionquant.tp=0 +ionquant.uniqueness=0 +ionquant.use-labeling=false +ionquant.use-lfq=true +ionquant.writeindex=0 +msbooster.predict-rt=true +msbooster.predict-spectra=true +msbooster.run-msbooster=true +msbooster.use-correlated-features=false +msfragger.Y_type_masses= +msfragger.activation_types=all +msfragger.allowed_missed_cleavage_1=2 +msfragger.allowed_missed_cleavage_2=2 +msfragger.calibrate_mass=2 +msfragger.check_spectral_files=true +msfragger.clip_nTerm_M=true +msfragger.deisotope=1 +msfragger.delta_mass_exclude_ranges=(-1.5,3.5) +msfragger.deneutralloss=1 +msfragger.diagnostic_fragments= +msfragger.diagnostic_intensity_filter=0 +msfragger.digest_max_length=50 +msfragger.digest_min_length=8 +msfragger.fragment_ion_series=b,y +msfragger.fragment_mass_tolerance=20 +msfragger.fragment_mass_units=1 +msfragger.group_variable=0 +msfragger.intensity_transform=0 +msfragger.ion_series_definitions= +msfragger.isotope_error=-1/0/1/2/3 +msfragger.labile_search_mode=off +msfragger.localize_delta_mass=false +msfragger.mass_diff_to_variable_mod=0 +msfragger.mass_offsets=0 +msfragger.mass_offsets_detailed= +msfragger.max_fragment_charge=2 +msfragger.max_variable_mods_combinations=5000 +msfragger.max_variable_mods_per_peptide=3 +msfragger.min_fragments_modelling=2 +msfragger.min_matched_fragments=4 +msfragger.min_sequence_matches=2 +msfragger.minimum_peaks=15 +msfragger.minimum_ratio=0.01 +msfragger.misc.fragger.clear-mz-hi=131.5 +msfragger.misc.fragger.clear-mz-lo=125.5 +msfragger.misc.fragger.digest-mass-hi=5000 +msfragger.misc.fragger.digest-mass-lo=200 +msfragger.misc.fragger.enzyme-dropdown-1=stricttrypsin +msfragger.misc.fragger.enzyme-dropdown-2=null +msfragger.misc.fragger.precursor-charge-hi=4 +msfragger.misc.fragger.precursor-charge-lo=1 +msfragger.misc.fragger.remove-precursor-range-hi=1.5 +msfragger.misc.fragger.remove-precursor-range-lo=-1.5 +msfragger.misc.slice-db=1 +msfragger.num_enzyme_termini=2 +msfragger.output_format=pepXML_pin +msfragger.output_max_expect=50 +msfragger.output_report_topN=1 +msfragger.output_report_topN_dda_plus=5 +msfragger.output_report_topN_dia1=5 +msfragger.override_charge=false +msfragger.precursor_mass_lower=-20 +msfragger.precursor_mass_mode=selected +msfragger.precursor_mass_units=1 +msfragger.precursor_mass_upper=20 +msfragger.precursor_true_tolerance=20 +msfragger.precursor_true_units=1 +msfragger.remainder_fragment_masses=-18.01056 79.96633 +msfragger.remove_precursor_peak=1 +msfragger.report_alternative_proteins=true +msfragger.require_precursor=true +msfragger.restrict_deltamass_to=all +msfragger.reuse_dia_fragment_peaks=false +msfragger.run-msfragger=true +msfragger.search_enzyme_cut_1=KR +msfragger.search_enzyme_cut_2= +msfragger.search_enzyme_name_1=stricttrypsin +msfragger.search_enzyme_name_2=null +msfragger.search_enzyme_nocut_1= +msfragger.search_enzyme_nocut_2= +msfragger.search_enzyme_sense_1=C +msfragger.search_enzyme_sense_2=C +msfragger.table.fix-mods=0.0,C-Term Peptide,true,-1; 0.0,N-Term Peptide,true,-1; 0.0,C-Term Protein,true,-1; 0.0,N-Term Protein,true,-1; 0.0,G (glycine),true,-1; 0.0,A (alanine),true,-1; 0.0,S (serine),true,-1; 0.0,P (proline),true,-1; 0.0,V (valine),true,-1; 0.0,T (threonine),true,-1; 57.02146,C (cysteine),true,-1; 0.0,L (leucine),true,-1; 0.0,I (isoleucine),true,-1; 0.0,N (asparagine),true,-1; 0.0,D (aspartic acid),true,-1; 0.0,Q (glutamine),true,-1; 229.16293,K (lysine),true,-1; 0.0,E (glutamic acid),true,-1; 0.0,M (methionine),true,-1; 0.0,H (histidine),true,-1; 0.0,F (phenylalanine),true,-1; 0.0,R (arginine),true,-1; 0.0,Y (tyrosine),true,-1; 0.0,W (tryptophan),true,-1; 0.0,B ,true,-1; 0.0,J,true,-1; 0.0,O,true,-1; 0.0,U,true,-1; 0.0,X,true,-1; 0.0,Z,true,-1 +msfragger.table.var-mods=15.9949,M,true,3; 42.0106,[^,true,1; 79.96633,STY,false,3; -17.0265,nQnC,false,1; -18.0106,nE,false,1; 229.16293,n^,true,1; 229.16293,S,true,1; 0.0,site_08,false,1; 0.0,site_09,false,1; 0.0,site_10,false,1; 0.0,site_11,false,1; 0.0,site_12,false,1; 0.0,site_13,false,1; 0.0,site_14,false,1; 0.0,site_15,false,1; 0.0,site_16,false,1 +msfragger.track_zero_topN=0 +msfragger.use_all_mods_in_first_search=false +msfragger.use_detailed_offsets=false +msfragger.use_topN_peaks=150 +msfragger.write_calibrated_mzml=false +msfragger.write_uncalibrated_mgf=false +msfragger.zero_bin_accept_expect=0 +msfragger.zero_bin_mult_expect=1 +opair.activation1=HCD +opair.activation2=ETD +opair.filterOxonium=true +opair.glyco_db= +opair.max_glycans=4 +opair.max_isotope_error=2 +opair.min_isotope_error=0 +opair.ms1_tol=20 +opair.ms2_tol=20 +opair.oxonium_filtering_file= +opair.oxonium_minimum_intensity=0.05 +opair.reverse_scan_order=false +opair.run-opair=false +opair.single_scan_type=false +peptide-prophet.cmd-opts=--decoyprobs --ppm --accmass --nonparam --expectscore +peptide-prophet.combine-pepxml=false +peptide-prophet.run-peptide-prophet=false +percolator.cmd-opts=--only-psms --no-terminate --post-processing-tdc +percolator.keep-tsv-files=false +percolator.min-prob=0.5 +percolator.run-percolator=true +phi-report.dont-use-prot-proph-file=false +phi-report.filter=--sequential --picked --psm 0.05 --pep 0.05 --prot 0.05 +phi-report.pep-level-summary=false +phi-report.philosoher-msstats=false +phi-report.print-decoys=false +phi-report.prot-level-summary=false +phi-report.remove-contaminants=false +phi-report.run-report=true +protein-prophet.cmd-opts=--maxppmdiff 2000000 --minprob 0.5 +protein-prophet.run-protein-prophet=true +ptmprophet.cmdline=true +ptmprophet.run-ptmprophet=false +ptmshepherd.adv_params=false +ptmshepherd.annotation-common=true +ptmshepherd.annotation-custom=false +ptmshepherd.annotation-glyco=false +ptmshepherd.annotation-unimod=false +ptmshepherd.annotation_file= +ptmshepherd.annotation_tol=0.01 +ptmshepherd.cap_y_ions= +ptmshepherd.decoy_type=1 +ptmshepherd.diag_ions= +ptmshepherd.diagmine_diagMinFoldChange=3.0 +ptmshepherd.diagmine_diagMinSpecDiff=00.2 +ptmshepherd.diagmine_fragMinFoldChange=3.0 +ptmshepherd.diagmine_fragMinPropensity=00.1 +ptmshepherd.diagmine_fragMinSpecDiff=00.1 +ptmshepherd.diagmine_minIonsPerSpec=2 +ptmshepherd.diagmine_minPeps=25 +ptmshepherd.diagmine_pepMinFoldChange=3.0 +ptmshepherd.diagmine_pepMinSpecDiff=00.2 +ptmshepherd.glyco_adducts= +ptmshepherd.glyco_fdr=5.00 +ptmshepherd.glyco_isotope_max=3 +ptmshepherd.glyco_isotope_min=-1 +ptmshepherd.glyco_ppm_tol=50 +ptmshepherd.glycodatabase= +ptmshepherd.histo_smoothbins=2 +ptmshepherd.iontype_a=false +ptmshepherd.iontype_b=true +ptmshepherd.iontype_c=false +ptmshepherd.iontype_x=false +ptmshepherd.iontype_y=true +ptmshepherd.iontype_z=false +ptmshepherd.localization_allowed_res= +ptmshepherd.localization_background=4 +ptmshepherd.max_adducts=0 +ptmshepherd.n_glyco=true +ptmshepherd.normalization-psms=true +ptmshepherd.normalization-scans=false +ptmshepherd.output_extended=false +ptmshepherd.peakpicking_mass_units=0 +ptmshepherd.peakpicking_minPsm=10 +ptmshepherd.peakpicking_promRatio=0.3 +ptmshepherd.peakpicking_width=0.002 +ptmshepherd.precursor_mass_units=0 +ptmshepherd.precursor_tol=0.01 +ptmshepherd.print_decoys=false +ptmshepherd.prob_dhexOx=2,0.5,0.1 +ptmshepherd.prob_dhexY=2,0.5 +ptmshepherd.prob_neuacOx=2,0.05,0.2 +ptmshepherd.prob_neugcOx=2,0.05,0.2 +ptmshepherd.prob_phosphoOx=2,0.05,0.2 +ptmshepherd.prob_regY=5,0.5 +ptmshepherd.prob_sulfoOx=2,0.05,0.2 +ptmshepherd.remainder_masses= +ptmshepherd.remove_glycan_delta_mass=true +ptmshepherd.run-shepherd=false +ptmshepherd.run_diagextract_mode=false +ptmshepherd.run_diagmine_mode=false +ptmshepherd.run_glyco_mode=false +ptmshepherd.spectra_maxfragcharge=2 +ptmshepherd.spectra_ppmtol=20 +ptmshepherd.varmod_masses= +quantitation.run-label-free-quant=false +run-psm-validation=true +run-validation-tab=true +saintexpress.fragpipe.cmd-opts= +saintexpress.max-replicates=3 +saintexpress.run-saint-express=true +saintexpress.virtual-controls=100 +speclibgen.easypqp.extras.max_delta_ppm=15 +speclibgen.easypqp.extras.max_delta_unimod=0.02 +speclibgen.easypqp.extras.rt_lowess_fraction=0 +speclibgen.easypqp.fragment.a=false +speclibgen.easypqp.fragment.b=true +speclibgen.easypqp.fragment.c=false +speclibgen.easypqp.fragment.x=false +speclibgen.easypqp.fragment.y=true +speclibgen.easypqp.fragment.z=false +speclibgen.easypqp.im-cal=Automatic selection of a run as reference IM +speclibgen.easypqp.neutral_loss=false +speclibgen.easypqp.rt-cal=noiRT +speclibgen.easypqp.select-file.text= +speclibgen.easypqp.select-im-file.text= +speclibgen.keep-intermediate-files=false +speclibgen.run-speclibgen=false +tab-run.delete_calibrated_mzml=false +tab-run.delete_temp_files=false +tab-run.sub_mzml_prob_threshold=0.5 +tab-run.write_sub_mzml=false +tmtintegrator.add_Ref=-1 +tmtintegrator.aggregation_method=0 +tmtintegrator.allow_overlabel=true +tmtintegrator.allow_unlabeled=false +tmtintegrator.best_psm=true +tmtintegrator.channel_num=TMT-11 +tmtintegrator.extraction_tool=IonQuant +tmtintegrator.glyco_qval=-1 +tmtintegrator.groupby=-1 +tmtintegrator.log2transformed=true +tmtintegrator.max_pep_prob_thres=0.9 +tmtintegrator.min_ntt=0 +tmtintegrator.min_pep_prob=0.9 +tmtintegrator.min_percent=0.05 +tmtintegrator.min_purity=0.5 +tmtintegrator.min_site_prob=-1 +tmtintegrator.mod_tag=none +tmtintegrator.ms1_int=true +tmtintegrator.outlier_removal=true +tmtintegrator.print_RefInt=false +tmtintegrator.prot_exclude=none +tmtintegrator.prot_norm=1 +tmtintegrator.psm_norm=false +tmtintegrator.quant_level=2 +tmtintegrator.ref_tag=Bridge +tmtintegrator.run-tmtintegrator=true +tmtintegrator.tolerance=20 +tmtintegrator.top3_pep=true +tmtintegrator.unique_gene=0 +tmtintegrator.unique_pep=false +tmtintegrator.use_glycan_composition=false +workflow.description=

Basic TMT 10-plex workflow, with identification and quantification from high mass accuracy MS2. Met oxidation, protein N-term Acetyl, n-term TMT, and TMT on S ("overlabeling") are specified as variable modifications. TMT-Integrator with virtual reference approach, median-centering normalization, data summarization at the gene level. If a reference/bridge sample is available, specify the corresponding channel/sample name tag in the annotation file(s) and in TMT-Integrator tab.

+workflow.input.data-type.im-ms=false +workflow.input.data-type.regular-ms=true +workflow.misc.save-sdrf=true +workflow.saved-with-ver=22.0 diff --git a/tumor-enriched-splicing/input/PDC000180filesmanifest.fp-manifest b/tumor-enriched-splicing/input/PDC000180filesmanifest.fp-manifest new file mode 100644 index 0000000..8bc0a4a --- /dev/null +++ b/tumor-enriched-splicing/input/PDC000180filesmanifest.fp-manifest @@ -0,0 +1,276 @@ +/home/rstudio/fragpipe/tumor-enriched-splicing/tmp/01CBTTC_PBT_Proteome_HMS_20180626/NCI-11plex-1-F12-f10283.mzML NCI_11plex_1_F12_f10283 DDA +/home/rstudio/fragpipe/tumor-enriched-splicing/tmp/01CBTTC_PBT_Proteome_HMS_20180626/NCI-11plex-1-F8-f10279.mzML NCI_11plex_1_F8_f10279 DDA +/home/rstudio/fragpipe/tumor-enriched-splicing/tmp/01CBTTC_PBT_Proteome_HMS_20180626/NCI-11plex-1-F11-f10282.mzML NCI_11plex_1_F11_f10282 DDA +/home/rstudio/fragpipe/tumor-enriched-splicing/tmp/01CBTTC_PBT_Proteome_HMS_20180626/NCI-11plex-1-F6-f10277.mzML NCI_11plex_1_F6_f10277 DDA 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NCI_11Plex_18_F12_f10527 DDA +/home/rstudio/fragpipe/tumor-enriched-splicing/tmp/18CBTTC_PBT_Proteome_HMS_20180626/NCI-11Plex-18-F7-f10522.mzML NCI_11Plex_18_F7_f10522 DDA +/home/rstudio/fragpipe/tumor-enriched-splicing/tmp/18CBTTC_PBT_Proteome_HMS_20180626/NCI-11Plex-18-F4-f10519.mzML NCI_11Plex_18_F4_f10519 DDA +/home/rstudio/fragpipe/tumor-enriched-splicing/tmp/19CBTTC_PBT_Proteome_HMS_20180626/NCI-11Plex-19-F7-f10550.mzML NCI_11Plex_19_F7_f10550 DDA +/home/rstudio/fragpipe/tumor-enriched-splicing/tmp/19CBTTC_PBT_Proteome_HMS_20180626/NCI-11Plex-19-F10-f10553.mzML NCI_11Plex_19_F10_f10553 DDA +/home/rstudio/fragpipe/tumor-enriched-splicing/tmp/19CBTTC_PBT_Proteome_HMS_20180626/NCI-11Plex-19-F11-f10554.mzML NCI_11Plex_19_F11_f10554 DDA +/home/rstudio/fragpipe/tumor-enriched-splicing/tmp/19CBTTC_PBT_Proteome_HMS_20180626/NCI-11Plex-19-F3-f10546.mzML NCI_11Plex_19_F3_f10546 DDA 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NCI_11Plex_20_F10_f10567 DDA +/home/rstudio/fragpipe/tumor-enriched-splicing/tmp/20CBTTC_PBT_Proteome_HMS_20180626/NCI-11Plex-20-F5-f10562.mzML NCI_11Plex_20_F5_f10562 DDA +/home/rstudio/fragpipe/tumor-enriched-splicing/tmp/20CBTTC_PBT_Proteome_HMS_20180626/NCI-11Plex-20-F9-f10566.mzML NCI_11Plex_20_F9_f10566 DDA +/home/rstudio/fragpipe/tumor-enriched-splicing/tmp/20CBTTC_PBT_Proteome_HMS_20180626/NCI-11Plex-20-F11-f10568.mzML NCI_11Plex_20_F11_f10568 DDA +/home/rstudio/fragpipe/tumor-enriched-splicing/tmp/20CBTTC_PBT_Proteome_HMS_20180626/NCI-11Plex-20-F7-f10564.mzML NCI_11Plex_20_F7_f10564 DDA +/home/rstudio/fragpipe/tumor-enriched-splicing/tmp/20CBTTC_PBT_Proteome_HMS_20180626/NCI-11Plex-20-F4-f10561.mzML NCI_11Plex_20_F4_f10561 DDA +/home/rstudio/fragpipe/tumor-enriched-splicing/tmp/20CBTTC_PBT_Proteome_HMS_20180626/NCI-11Plex-20-F1-f10558.mzML NCI_11Plex_20_F1_f10558 DDA 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DDA +/home/rstudio/fragpipe/tumor-enriched-splicing/tmp/21CBTTC_PBT_Proteome_HMS_20180626/NCI-11Plex-21-F12-m17287.mzML NCI_11Plex_21_F12_m17287 DDA +/home/rstudio/fragpipe/tumor-enriched-splicing/tmp/21CBTTC_PBT_Proteome_HMS_20180626/NCI-11Plex-21-F8-m17283.mzML NCI_11Plex_21_F8_m17283 DDA +/home/rstudio/fragpipe/tumor-enriched-splicing/tmp/21CBTTC_PBT_Proteome_HMS_20180626/NCI-11Plex-21-F11-m17286.mzML NCI_11Plex_21_F11_m17286 DDA +/home/rstudio/fragpipe/tumor-enriched-splicing/tmp/21CBTTC_PBT_Proteome_HMS_20180626/NCI-11Plex-21-F4-m17279.mzML NCI_11Plex_21_F4_m17279 DDA +/home/rstudio/fragpipe/tumor-enriched-splicing/tmp/21CBTTC_PBT_Proteome_HMS_20180626/NCI-11Plex-21-F2-m17277.mzML NCI_11Plex_21_F2_m17277 DDA +/home/rstudio/fragpipe/tumor-enriched-splicing/tmp/22CBTTC_PBT_Proteome_HMS_20180626/NCI-11Plex-22-F1-m17289.mzML NCI_11Plex_22_F1_m17289 DDA +/home/rstudio/fragpipe/tumor-enriched-splicing/tmp/22CBTTC_PBT_Proteome_HMS_20180626/NCI-11Plex-22-F4-m17292.mzML NCI_11Plex_22_F4_m17292 DDA +/home/rstudio/fragpipe/tumor-enriched-splicing/tmp/22CBTTC_PBT_Proteome_HMS_20180626/NCI-11Plex-22-F3-m17291.mzML NCI_11Plex_22_F3_m17291 DDA +/home/rstudio/fragpipe/tumor-enriched-splicing/tmp/22CBTTC_PBT_Proteome_HMS_20180626/NCI-11Plex-22-F11-m17299.mzML NCI_11Plex_22_F11_m17299 DDA +/home/rstudio/fragpipe/tumor-enriched-splicing/tmp/22CBTTC_PBT_Proteome_HMS_20180626/NCI-11Plex-22-F9-m17297.mzML NCI_11Plex_22_F9_m17297 DDA +/home/rstudio/fragpipe/tumor-enriched-splicing/tmp/22CBTTC_PBT_Proteome_HMS_20180626/NCI-11Plex-22-F10-m17298.mzML NCI_11Plex_22_F10_m17298 DDA +/home/rstudio/fragpipe/tumor-enriched-splicing/tmp/22CBTTC_PBT_Proteome_HMS_20180626/NCI-11Plex-22-F5-m17293.mzML NCI_11Plex_22_F5_m17293 DDA +/home/rstudio/fragpipe/tumor-enriched-splicing/tmp/22CBTTC_PBT_Proteome_HMS_20180626/NCI-11Plex-22-F2-m17290.mzML NCI_11Plex_22_F2_m17290 DDA 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DDA +/home/rstudio/fragpipe/tumor-enriched-splicing/tmp/23CBTTC_PBT_Proteome_HMS_20180626/NCI-11Plex-23-F3-m17317.mzML NCI_11Plex_23_F3_m17317 DDA +/home/rstudio/fragpipe/tumor-enriched-splicing/tmp/23CBTTC_PBT_Proteome_HMS_20180626/NCI-11Plex-23-F2-m17316.mzML NCI_11Plex_23_F2_m17316 DDA +/home/rstudio/fragpipe/tumor-enriched-splicing/tmp/23CBTTC_PBT_Proteome_HMS_20180626/NCI-11Plex-23-F1-m17315.mzML NCI_11Plex_23_F1_m17315 DDA +/home/rstudio/fragpipe/tumor-enriched-splicing/tmp/23CBTTC_PBT_Proteome_HMS_20180626/NCI-11Plex-23-F4-m17318.mzML NCI_11Plex_23_F4_m17318 DDA +/home/rstudio/fragpipe/tumor-enriched-splicing/tmp/23CBTTC_PBT_Proteome_HMS_20180626/NCI-11Plex-23-F11-m17325.mzML NCI_11Plex_23_F11_m17325 DDA +/home/rstudio/fragpipe/tumor-enriched-splicing/tmp/23CBTTC_PBT_Proteome_HMS_20180626/NCI-11Plex-23-F10-m17324.mzML NCI_11Plex_23_F10_m17324 DDA +/home/rstudio/fragpipe/tumor-enriched-splicing/tmp/23CBTTC_PBT_Proteome_HMS_20180626/NCI-11Plex-23-F5-m17319.mzML NCI_11Plex_23_F5_m17319 DDA +/home/rstudio/fragpipe/tumor-enriched-splicing/tmp/23CBTTC_PBT_Proteome_HMS_20180626/NCI-11Plex-23-F12-m17326.mzML NCI_11Plex_23_F12_m17326 DDA +/home/rstudio/fragpipe/tumor-enriched-splicing/tmp/23CBTTC_PBT_Proteome_HMS_20180626/NCI-11Plex-23-F6-m17320.mzML NCI_11Plex_23_F6_m17320 DDA diff --git a/tumor-enriched-splicing/run_fragpipe.sh b/tumor-enriched-splicing/run_fragpipe.sh new file mode 100644 index 0000000..716dc5c --- /dev/null +++ b/tumor-enriched-splicing/run_fragpipe.sh @@ -0,0 +1,118 @@ +#!/bin/bash + +set -e +set -o pipefail + +# Set tools and input directories +input_dir=input +tools_dir=/fragpipe_bin/fragPipe-22.0/fragpipe/tools/ + +# Error out if no custom.fasta is provided +if [ ! -e "$input_dir/custom.fasta" ]; then + echo "Error: must supply a custom.fasta file in the input directory ($input_dir)" + exit 1 +fi + +echo "Adding decoys and contaminants to FASTA files.." +cd $input_dir + +# Initialize the Philosopher workspace +$tools_dir/Philosopher/philosopher-v5.1.1 workspace --init + +Uniprot_canonical=/home/rstudio/fragpipe/refs/UP000005640_9606.fasta.gz + +# Add decoys and contaminants to the custom FASTA file +gunzip -c $Uniprot_canonical | $tools_dir/Philosopher/philosopher-v5.1.1 database --custom custom.fasta --add /dev/stdin --contam +#$tools_dir/Philosopher/philosopher-v5.1.1 database --custom custom.fasta --contam + +#mv *decoys-contam-custom.fasta.fas decoys-contam-custom-canonical.fasta +# Remove canonica peptides annotated to genes with TESEs +grep "^>" custom.fasta | awk -F'[:_]' '{print "GN=" $2 " "}' | sort -u > gene_symbols.txt + +awk 'BEGIN{while((getline k < "gene_symbols.txt") > 0){a[k]=1}} + /^>/{ + keep=1 + for (g in a) { + if (index($0,g)) {keep=0; break} + } + } + keep' *decoys-contam-custom.fasta.fas > decoys-contam-custom-canonical.fasta + +# Clean intermediate files +$tools_dir/Philosopher/philosopher-v5.1.1 workspace --clean + +# Return to the previous directory +cd - + +# define directory to mount cavatica project; create if it does not exist +data_dir=/home/rstudio/fragpipe/tumor-enriched-splicing/cavatica-data + +if [ ! -d $data_dir ]; then + mkdir -p $data_dir + echo "Creating cavatica-data directory to mount cavatica project..." +fi + +# set cavatica directory +cavatica_dir=/home/rstudio/fragpipe/tumor-enriched-splicing/cavatica-data/projects/harenzaj/proteomics + +if [ ! -d $cavatica_dir ]; then + echo "Error: path to cavatica project directory does not exist. Attempting to mount project with sbfs mount..." + + sbfs mount --profile default --project harenzaj/proteomics cavatica-data + +fi + +# check if tmp directory exists, and create it if not +tmp_dir=(/home/rstudio/fragpipe/tumor-enriched-splicing/tmp) + +if [ ! -d $tmp_dir ]; then + mkdir -p $tmp_dir + echo "Temporary directory does not exist, creating..." +fi + +# clear contents of tmp directory, if not already empty +rm -rf $tmp_dir/* + +# copy mzML files to temporary directory, maintaining directory structure +echo "copying mzML files to temporary directory..." + +if ! [[ " $* " == *" --run_subset "* ]]; then + +cp -R $cavatica_dir/*CBTTC_PBT_Proteome* $tmp_dir + +else + +cp -R $cavatica_dir/01CBTTC_PBT_Proteome* $tmp_dir + +fi + +# unzip mzML files, force overwrite if files exist +echo "unzipping mzML files..." +gunzip -f $tmp_dir/*CBTTC_PBT_Proteome*/*mzML.gz + +# set workflow, manifest, results and tools directories +echo "Starting Fragpipe run..." + +wf=/home/rstudio/fragpipe/tumor-enriched-splicing/input/PDC000180customworkflow.workflow +manifest=/home/rstudio/fragpipe/tumor-enriched-splicing/input/PDC000180filesmanifest.fp-manifest + +if [[ " $* " == *" --run_subset "* ]]; then + +grep "01CBTTC_" $manifest > /home/rstudio/fragpipe/tumor-enriched-splicing/input/PDC000180filesmanifest.fp-manifest_sub +manifest=/home/rstudio/fragpipe/tumor-enriched-splicing/input/PDC000180filesmanifest.fp-manifest_sub + +fi + +res_dir=/home/rstudio/fragpipe/tumor-enriched-splicing/results-cptac-filtered-canon + +if [ ! -d $res_dir ]; then + mkdir -p $res_dir + echo "Creating results directory..." +fi + +tools_dir=/fragpipe_bin/fragPipe-22.0/fragpipe/tools/ + +/fragpipe_bin/fragPipe-22.0/fragpipe/bin/fragpipe --headless --workflow $wf --manifest $manifest --workdir $res_dir --config-tools-folder $tools_dir + +# clear tmp directory +rm -rf $tmp_dir/*