Annotate Misprime 6

Last updated: 2020-03-17

Checks: 7 0

Knit directory: Comparative_APA/analysis/

This reproducible R Markdown analysis was created with workflowr (version 1.6.0). The Checks tab describes the reproducibility checks that were applied when the results were created. The Past versions tab lists the development history.

R Markdown file: up-to-date

Great! Since the R Markdown file has been committed to the Git repository, you know the exact version of the code that produced these results.

Environment: empty

Great job! The global environment was empty. Objects defined in the global environment can affect the analysis in your R Markdown file in unknown ways. For reproduciblity it’s best to always run the code in an empty environment.

Seed: set.seed(20190902)

The command set.seed(20190902) was run prior to running the code in the R Markdown file. Setting a seed ensures that any results that rely on randomness, e.g. subsampling or permutations, are reproducible.

Session information: recorded

Great job! Recording the operating system, R version, and package versions is critical for reproducibility.

Cache: none

Nice! There were no cached chunks for this analysis, so you can be confident that you successfully produced the results during this run.

File paths: relative

Great job! Using relative paths to the files within your workflowr project makes it easier to run your code on other machines.

Repository version: bec8ae9

Great! You are using Git for version control. Tracking code development and connecting the code version to the results is critical for reproducibility. The version displayed above was the version of the Git repository at the time these results were generated.

Note that you need to be careful to ensure that all relevant files for the analysis have been committed to Git prior to generating the results (you can use wflow_publish or wflow_git_commit). workflowr only checks the R Markdown file, but you know if there are other scripts or data files that it depends on. Below is the status of the Git repository when the results were generated:


Ignored files:
    Ignored:    .DS_Store
    Ignored:    .Rhistory
    Ignored:    .Rproj.user/
    Ignored:    code/chimp_log/
    Ignored:    code/human_log/
    Ignored:    data/.DS_Store
    Ignored:    data/mediation_prot/
    Ignored:    data/metadata_HCpanel.txt.sb-a5794dd2-i594qs/
    Ignored:    output/.DS_Store

Untracked files:
    Untracked:  ._.DS_Store
    Untracked:  Chimp/
    Untracked:  Human/
    Untracked:  analysis/CrossChimpThreePrime.Rmd
    Untracked:  analysis/DiffTransProtvsExpression.Rmd
    Untracked:  analysis/DiffUsedUTR.Rmd
    Untracked:  analysis/GvizPlots.Rmd
    Untracked:  analysis/HandC.TvN
    Untracked:  analysis/PhenotypeOverlap10.Rmd
    Untracked:  analysis/annotationBias.Rmd
    Untracked:  analysis/assessReadQual.Rmd
    Untracked:  analysis/diffExpressionPantro6.Rmd
    Untracked:  analysis/orthoexonAnno.Rmd
    Untracked:  analysis/pol2.Rmd
    Untracked:  code/._ClassifyLeafviz.sh
    Untracked:  code/._Config_chimp.yaml
    Untracked:  code/._Config_chimp_full.yaml
    Untracked:  code/._Config_human.yaml
    Untracked:  code/._ConvertJunc2Bed.sh
    Untracked:  code/._CountNucleotides.py
    Untracked:  code/._CrossMapChimpRNA.sh
    Untracked:  code/._CrossMapThreeprime.sh
    Untracked:  code/._DiffSplice.sh
    Untracked:  code/._DiffSplicePlots.sh
    Untracked:  code/._DiffSplicePlots_gencode.sh
    Untracked:  code/._DiffSplice_gencode.sh
    Untracked:  code/._DiffSplice_removebad.sh
    Untracked:  code/._FindIntronForDomPAS.sh
    Untracked:  code/._FindIntronForDomPAS_DF.sh
    Untracked:  code/._GetMAPQscore.py
    Untracked:  code/._GetSecondaryMap.py
    Untracked:  code/._Lift5perPAS.sh
    Untracked:  code/._LiftFinalChimpJunc2Human.sh
    Untracked:  code/._LiftOrthoPAS2chimp.sh
    Untracked:  code/._MapBadSamples.sh
    Untracked:  code/._PAS_ATTAAA.sh
    Untracked:  code/._PAS_ATTAAA_df.sh
    Untracked:  code/._PAS_seqExpanded.sh
    Untracked:  code/._PASsequences.sh
    Untracked:  code/._PASsequences_DF.sh
    Untracked:  code/._PlotNuclearUsagebySpecies.R
    Untracked:  code/._PlotNuclearUsagebySpecies_DF.R
    Untracked:  code/._QuantMergedClusters.sh
    Untracked:  code/._RNATranscriptDTplot.sh
    Untracked:  code/._ReverseLiftFilter.R
    Untracked:  code/._RunFixLeafCluster.sh
    Untracked:  code/._RunNegMCMediation.sh
    Untracked:  code/._RunNegMCMediationDF.sh
    Untracked:  code/._RunPosMCMediationDF.err
    Untracked:  code/._RunPosMCMediationDF.sh
    Untracked:  code/._SAF2Bed.py
    Untracked:  code/._Snakefile
    Untracked:  code/._SnakefilePAS
    Untracked:  code/._SnakefilePASfilt
    Untracked:  code/._SortIndexBadSamples.sh
    Untracked:  code/._assignPeak2Intronicregion
    Untracked:  code/._assignPeak2Intronicregion.sh
    Untracked:  code/._bed215upbed.py
    Untracked:  code/._bed2SAF_gen.py
    Untracked:  code/._buildIndecpantro5
    Untracked:  code/._buildIndecpantro5.sh
    Untracked:  code/._buildLeafviz.sh
    Untracked:  code/._buildLeafviz_leadAnno.sh
    Untracked:  code/._buildStarIndex.sh
    Untracked:  code/._chimpChromprder.sh
    Untracked:  code/._chooseSignalSite.py
    Untracked:  code/._cleanbed2saf.py
    Untracked:  code/._cluster.json
    Untracked:  code/._cluster2bed.py
    Untracked:  code/._clusterLiftReverse.sh
    Untracked:  code/._clusterLiftReverse_removebad.sh
    Untracked:  code/._clusterLiftprimary.sh
    Untracked:  code/._clusterLiftprimary_removebad.sh
    Untracked:  code/._converBam2Junc.sh
    Untracked:  code/._converBam2Junc_removeBad.sh
    Untracked:  code/._extraSnakefiltpas
    Untracked:  code/._extractPhyloReg.py
    Untracked:  code/._extractPhyloRegGene.py
    Untracked:  code/._extractPhylopReg200down.py
    Untracked:  code/._extractPhylopReg200up.py
    Untracked:  code/._filter5percPAS.py
    Untracked:  code/._filterNumChroms.py
    Untracked:  code/._filterPASforMP.py
    Untracked:  code/._filterPostLift.py
    Untracked:  code/._fixExonFC.py
    Untracked:  code/._fixLeafCluster.py
    Untracked:  code/._fixLiftedJunc.py
    Untracked:  code/._fixUTRexonanno.py
    Untracked:  code/._formathg38Anno.py
    Untracked:  code/._formatpantro6Anno.py
    Untracked:  code/._getRNAseqMapStats.sh
    Untracked:  code/._hg19MapStats.sh
    Untracked:  code/._humanChromorder.sh
    Untracked:  code/._intersectLiftedPAS.sh
    Untracked:  code/._liftJunctionFiles.sh
    Untracked:  code/._liftPAS19to38.sh
    Untracked:  code/._liftedchimpJunc2human.sh
    Untracked:  code/._makeNuclearDapaplots.sh
    Untracked:  code/._makeNuclearDapaplots_DF.sh
    Untracked:  code/._makeSamplyGroupsHuman_TvN.py
    Untracked:  code/._mapRNAseqhg19.sh
    Untracked:  code/._mapRNAseqhg19_newPipeline.sh
    Untracked:  code/._maphg19.sh
    Untracked:  code/._maphg19_subjunc.sh
    Untracked:  code/._mediation_test.R
    Untracked:  code/._mergeChimp3prime_inhg38.sh
    Untracked:  code/._mergeandBWRNAseq.sh
    Untracked:  code/._mergedBam2BW.sh
    Untracked:  code/._nameClusters.py
    Untracked:  code/._negativeMediation_montecarlo.R
    Untracked:  code/._negativeMediation_montecarloDF.R
    Untracked:  code/._numMultimap.py
    Untracked:  code/._overlapapaQTLPAS.sh
    Untracked:  code/._parseHg38.py
    Untracked:  code/._postiveMediation_montecarlo_DF.R
    Untracked:  code/._prepareCleanLiftedFC_5perc4LC.py
    Untracked:  code/._prepareLeafvizAnno.sh
    Untracked:  code/._preparePAS4lift.py
    Untracked:  code/._primaryLift.sh
    Untracked:  code/._processhg38exons.py
    Untracked:  code/._quantJunc.sh
    Untracked:  code/._quantJunc_TEST.sh
    Untracked:  code/._quantJunc_removeBad.sh
    Untracked:  code/._quantMerged_seperatly.sh
    Untracked:  code/._recLiftchim2human.sh
    Untracked:  code/._revLiftPAShg38to19.sh
    Untracked:  code/._reverseLift.sh
    Untracked:  code/._runCheckReverseLift.sh
    Untracked:  code/._runChimpDiffIso.sh
    Untracked:  code/._runCountNucleotides.sh
    Untracked:  code/._runFilterNumChroms.sh
    Untracked:  code/._runHumanDiffIso.sh
    Untracked:  code/._runNuclearDiffIso_DF.sh
    Untracked:  code/._runNuclearDifffIso.sh
    Untracked:  code/._runTotalDiffIso.sh
    Untracked:  code/._run_chimpverifybam.sh
    Untracked:  code/._run_verifyBam.sh
    Untracked:  code/._snakemake.batch
    Untracked:  code/._snakemakePAS.batch
    Untracked:  code/._snakemakePASchimp.batch
    Untracked:  code/._snakemakePAShuman.batch
    Untracked:  code/._snakemake_chimp.batch
    Untracked:  code/._snakemake_human.batch
    Untracked:  code/._snakemakefiltPAS.batch
    Untracked:  code/._snakemakefiltPAS_chimp
    Untracked:  code/._snakemakefiltPAS_chimp.sh
    Untracked:  code/._snakemakefiltPAS_human.sh
    Untracked:  code/._spliceSite2Fasta.py
    Untracked:  code/._submit-snakemake-chimp.sh
    Untracked:  code/._submit-snakemake-human.sh
    Untracked:  code/._submit-snakemakePAS-chimp.sh
    Untracked:  code/._submit-snakemakePAS-human.sh
    Untracked:  code/._submit-snakemakefiltPAS-chimp.sh
    Untracked:  code/._submit-snakemakefiltPAS-human.sh
    Untracked:  code/._subset_diffisopheno_Nuclear_HvC.py
    Untracked:  code/._subset_diffisopheno_Nuclear_HvC_DF.py
    Untracked:  code/._subset_diffisopheno_Total_HvC.py
    Untracked:  code/._threeprimeOrthoFC.sh
    Untracked:  code/._transcriptDTplotsNuclear.sh
    Untracked:  code/._verifyBam4973.sh
    Untracked:  code/._verifyBam4973inHuman.sh
    Untracked:  code/._wrap_chimpverifybam.sh
    Untracked:  code/._wrap_verifyBam.sh
    Untracked:  code/._writeMergecode.py
    Untracked:  code/.snakemake/
    Untracked:  code/ClassifyLeafviz.sh
    Untracked:  code/Config_chimp.yaml
    Untracked:  code/Config_chimp_full.yaml
    Untracked:  code/Config_human.yaml
    Untracked:  code/ConvertJunc2Bed.err
    Untracked:  code/ConvertJunc2Bed.out
    Untracked:  code/ConvertJunc2Bed.sh
    Untracked:  code/CountNucleotides.py
    Untracked:  code/CrossMapChimpRNA.sh
    Untracked:  code/CrossMapThreeprime.sh
    Untracked:  code/CrossmapChimp3prime.err
    Untracked:  code/CrossmapChimp3prime.out
    Untracked:  code/CrossmapChimpRNA.err
    Untracked:  code/CrossmapChimpRNA.out
    Untracked:  code/DiffSplice.err
    Untracked:  code/DiffSplice.out
    Untracked:  code/DiffSplice.sh
    Untracked:  code/DiffSplicePlots.err
    Untracked:  code/DiffSplicePlots.out
    Untracked:  code/DiffSplicePlots.sh
    Untracked:  code/DiffSplicePlots_gencode.sh
    Untracked:  code/DiffSplice_gencode.sh
    Untracked:  code/DiffSplice_removebad.err
    Untracked:  code/DiffSplice_removebad.out
    Untracked:  code/DiffSplice_removebad.sh
    Untracked:  code/FilterReverseLift.err
    Untracked:  code/FilterReverseLift.out
    Untracked:  code/FindIntronForDomPAS.err
    Untracked:  code/FindIntronForDomPAS.out
    Untracked:  code/FindIntronForDomPAS.sh
    Untracked:  code/FindIntronForDomPAS_DF.sh
    Untracked:  code/GencodeDiffSplice.err
    Untracked:  code/GencodeDiffSplice.out
    Untracked:  code/GetMAPQscore.py
    Untracked:  code/GetSecondaryMap.py
    Untracked:  code/HchromOrder.err
    Untracked:  code/HchromOrder.out
    Untracked:  code/JunctionLift.err
    Untracked:  code/JunctionLift.out
    Untracked:  code/JunctionLiftFinalChimp.err
    Untracked:  code/JunctionLiftFinalChimp.out
    Untracked:  code/Lift5perPAS.sh
    Untracked:  code/Lift5perPASbed.err
    Untracked:  code/Lift5perPASbed.out
    Untracked:  code/LiftClustersFirst.err
    Untracked:  code/LiftClustersFirst.out
    Untracked:  code/LiftClustersFirst_remove.err
    Untracked:  code/LiftClustersFirst_remove.out
    Untracked:  code/LiftClustersSecond.err
    Untracked:  code/LiftClustersSecond.out
    Untracked:  code/LiftClustersSecond_remove.err
    Untracked:  code/LiftClustersSecond_remove.out
    Untracked:  code/LiftFinalChimpJunc2Human.sh
    Untracked:  code/LiftOrthoPAS2chimp.sh
    Untracked:  code/LiftorthoPAS.err
    Untracked:  code/LiftorthoPASt.out
    Untracked:  code/Log.out
    Untracked:  code/MapBadSamples.err
    Untracked:  code/MapBadSamples.out
    Untracked:  code/MapBadSamples.sh
    Untracked:  code/MapStats.err
    Untracked:  code/MapStats.out
    Untracked:  code/MaxEntCode/
    Untracked:  code/MergeClusters.err
    Untracked:  code/MergeClusters.out
    Untracked:  code/MergeClusters.sh
    Untracked:  code/PAS_ATTAAA.err
    Untracked:  code/PAS_ATTAAA.out
    Untracked:  code/PAS_ATTAAA.sh
    Untracked:  code/PAS_ATTAAADF.err
    Untracked:  code/PAS_ATTAAADF.out
    Untracked:  code/PAS_ATTAAA_df.sh
    Untracked:  code/PAS_seqExpanded.sh
    Untracked:  code/PAS_sequence.err
    Untracked:  code/PAS_sequence.out
    Untracked:  code/PAS_sequenceDF.err
    Untracked:  code/PAS_sequenceDF.out
    Untracked:  code/PASexpanded_sequenceDF.err
    Untracked:  code/PASexpanded_sequenceDF.out
    Untracked:  code/PASsequences.sh
    Untracked:  code/PASsequences_DF.sh
    Untracked:  code/PlotNuclearUsagebySpecies.R
    Untracked:  code/PlotNuclearUsagebySpecies_DF.R
    Untracked:  code/QuantMergeClusters
    Untracked:  code/QuantMergeClusters.err
    Untracked:  code/QuantMergeClusters.out
    Untracked:  code/QuantMergedClusters.sh
    Untracked:  code/RNATranscriptDTplot.err
    Untracked:  code/RNATranscriptDTplot.out
    Untracked:  code/RNATranscriptDTplot.sh
    Untracked:  code/Rev_liftoverPAShg19to38.err
    Untracked:  code/Rev_liftoverPAShg19to38.out
    Untracked:  code/ReverseLiftFilter.R
    Untracked:  code/RunFixCluster.err
    Untracked:  code/RunFixCluster.out
    Untracked:  code/RunFixLeafCluster.sh
    Untracked:  code/RunNegMCMediation.err
    Untracked:  code/RunNegMCMediation.sh
    Untracked:  code/RunNegMCMediationDF.err
    Untracked:  code/RunNegMCMediationDF.out
    Untracked:  code/RunNegMCMediationDF.sh
    Untracked:  code/RunNegMCMediationr.out
    Untracked:  code/RunPosMCMediation.err
    Untracked:  code/RunPosMCMediation.sh
    Untracked:  code/RunPosMCMediationDF.err
    Untracked:  code/RunPosMCMediationDF.out
    Untracked:  code/RunPosMCMediationDF.sh
    Untracked:  code/RunPosMCMediationr.out
    Untracked:  code/SAF215upbed_gen.py
    Untracked:  code/SAF2Bed.py
    Untracked:  code/Snakefile
    Untracked:  code/SnakefilePAS
    Untracked:  code/SnakefilePASfilt
    Untracked:  code/SortIndexBadSamples.err
    Untracked:  code/SortIndexBadSamples.out
    Untracked:  code/SortIndexBadSamples.sh
    Untracked:  code/TotalTranscriptDTplot.err
    Untracked:  code/TotalTranscriptDTplot.out
    Untracked:  code/Upstream10Bases_general.py
    Untracked:  code/apaQTLsnake.err
    Untracked:  code/apaQTLsnake.out
    Untracked:  code/apaQTLsnakePAS.err
    Untracked:  code/apaQTLsnakePAS.out
    Untracked:  code/apaQTLsnakePAShuman.err
    Untracked:  code/assignPeak2Intronicregion.err
    Untracked:  code/assignPeak2Intronicregion.out
    Untracked:  code/assignPeak2Intronicregion.sh
    Untracked:  code/bam2junc.err
    Untracked:  code/bam2junc.out
    Untracked:  code/bam2junc_remove.err
    Untracked:  code/bam2junc_remove.out
    Untracked:  code/bed215upbed.py
    Untracked:  code/bed2SAF_gen.py
    Untracked:  code/bed2saf.py
    Untracked:  code/bg_to_cov.py
    Untracked:  code/buildIndecpantro5
    Untracked:  code/buildIndecpantro5.sh
    Untracked:  code/buildLeafviz.err
    Untracked:  code/buildLeafviz.out
    Untracked:  code/buildLeafviz.sh
    Untracked:  code/buildLeafviz_leadAnno.sh
    Untracked:  code/buildLeafviz_leafanno.err
    Untracked:  code/buildLeafviz_leafanno.out
    Untracked:  code/buildStarIndex.sh
    Untracked:  code/callPeaksYL.py
    Untracked:  code/chimpChromprder.sh
    Untracked:  code/chooseAnno2Bed.py
    Untracked:  code/chooseAnno2SAF.py
    Untracked:  code/chooseSignalSite.py
    Untracked:  code/chromOrder.err
    Untracked:  code/chromOrder.out
    Untracked:  code/classifyLeafviz.err
    Untracked:  code/classifyLeafviz.out
    Untracked:  code/cleanbed2saf.py
    Untracked:  code/cluster.json
    Untracked:  code/cluster2bed.py
    Untracked:  code/clusterLiftReverse.sh
    Untracked:  code/clusterLiftReverse_removebad.sh
    Untracked:  code/clusterLiftprimary.sh
    Untracked:  code/clusterLiftprimary_removebad.sh
    Untracked:  code/clusterPAS.json
    Untracked:  code/clusterfiltPAS.json
    Untracked:  code/comands2Mege.sh
    Untracked:  code/converBam2Junc.sh
    Untracked:  code/converBam2Junc_removeBad.sh
    Untracked:  code/convertNumeric.py
    Untracked:  code/environment.yaml
    Untracked:  code/extraSnakefiltpas
    Untracked:  code/extractPhyloReg.py
    Untracked:  code/extractPhyloRegGene.py
    Untracked:  code/extractPhylopReg200down.py
    Untracked:  code/extractPhylopReg200up.py
    Untracked:  code/filter5perc.R
    Untracked:  code/filter5percPAS.py
    Untracked:  code/filter5percPheno.py
    Untracked:  code/filterBamforMP.pysam2_gen.py
    Untracked:  code/filterJuncChroms.err
    Untracked:  code/filterJuncChroms.out
    Untracked:  code/filterMissprimingInNuc10_gen.py
    Untracked:  code/filterNumChroms.py
    Untracked:  code/filterPASforMP.py
    Untracked:  code/filterPostLift.py
    Untracked:  code/filterSAFforMP_gen.py
    Untracked:  code/filterSortBedbyCleanedBed_gen.R
    Untracked:  code/filterpeaks.py
    Untracked:  code/fixExonFC.py
    Untracked:  code/fixFChead.py
    Untracked:  code/fixFChead_bothfrac.py
    Untracked:  code/fixLeafCluster.py
    Untracked:  code/fixLiftedJunc.py
    Untracked:  code/fixUTRexonanno.py
    Untracked:  code/formathg38Anno.py
    Untracked:  code/generateStarIndex.err
    Untracked:  code/generateStarIndex.out
    Untracked:  code/generateStarIndexHuman.err
    Untracked:  code/generateStarIndexHuman.out
    Untracked:  code/getRNAseqMapStats.sh
    Untracked:  code/hg19MapStats.err
    Untracked:  code/hg19MapStats.out
    Untracked:  code/hg19MapStats.sh
    Untracked:  code/humanChromorder.sh
    Untracked:  code/humanFiles
    Untracked:  code/intersectAnno.err
    Untracked:  code/intersectAnno.out
    Untracked:  code/intersectAnnoExt.err
    Untracked:  code/intersectAnnoExt.out
    Untracked:  code/intersectLiftedPAS.sh
    Untracked:  code/leafcutter_merge_regtools_redo.py
    Untracked:  code/liftJunctionFiles.sh
    Untracked:  code/liftPAS19to38.sh
    Untracked:  code/liftoverPAShg19to38.err
    Untracked:  code/liftoverPAShg19to38.out
    Untracked:  code/log/
    Untracked:  code/make5percPeakbed.py
    Untracked:  code/makeFileID.py
    Untracked:  code/makeNuclearDapaplots.sh
    Untracked:  code/makeNuclearDapaplots_DF.sh
    Untracked:  code/makeNuclearPlots.err
    Untracked:  code/makeNuclearPlots.out
    Untracked:  code/makeNuclearPlotsDF.err
    Untracked:  code/makeNuclearPlotsDF.out
    Untracked:  code/makePheno.py
    Untracked:  code/makeSamplyGroupsChimp_TvN.py
    Untracked:  code/makeSamplyGroupsHuman_TvN.py
    Untracked:  code/mapRNAseqhg19.sh
    Untracked:  code/mapRNAseqhg19_newPipeline.sh
    Untracked:  code/maphg19.err
    Untracked:  code/maphg19.out
    Untracked:  code/maphg19.sh
    Untracked:  code/maphg19_new.err
    Untracked:  code/maphg19_new.out
    Untracked:  code/maphg19_sub.err
    Untracked:  code/maphg19_sub.out
    Untracked:  code/maphg19_subjunc.sh
    Untracked:  code/mediation_test.R
    Untracked:  code/merge.err
    Untracked:  code/mergeChimp3prime_inhg38.sh
    Untracked:  code/merge_leafcutter_clusters_redo.py
    Untracked:  code/mergeandBWRNAseq.sh
    Untracked:  code/mergeandsort_ChimpinHuman.err
    Untracked:  code/mergeandsort_ChimpinHuman.out
    Untracked:  code/mergedBam2BW.sh
    Untracked:  code/mergedbam2bw.err
    Untracked:  code/mergedbam2bw.out
    Untracked:  code/mergedbamRNAand2bw.err
    Untracked:  code/mergedbamRNAand2bw.out
    Untracked:  code/nameClusters.py
    Untracked:  code/namePeaks.py
    Untracked:  code/negativeMediation_montecarlo.R
    Untracked:  code/negativeMediation_montecarloDF.R
    Untracked:  code/nuclearTranscriptDTplot.err
    Untracked:  code/nuclearTranscriptDTplot.out
    Untracked:  code/numMultimap.py
    Untracked:  code/overlapPAS.err
    Untracked:  code/overlapPAS.out
    Untracked:  code/overlapapaQTLPAS.sh
    Untracked:  code/overlapapaQTLPAS_extended.sh
    Untracked:  code/overlapapaQTLPAS_samples.sh
    Untracked:  code/parseHg38.py
    Untracked:  code/peak2PAS.py
    Untracked:  code/pheno2countonly.R
    Untracked:  code/postiveMediation_montecarlo.R
    Untracked:  code/postiveMediation_montecarlo_DF.R
    Untracked:  code/prepareAnnoLeafviz.err
    Untracked:  code/prepareAnnoLeafviz.out
    Untracked:  code/prepareCleanLiftedFC_5perc4LC.py
    Untracked:  code/prepareLeafvizAnno.sh
    Untracked:  code/preparePAS4lift.py
    Untracked:  code/prepare_phenotype_table.py
    Untracked:  code/primaryLift.err
    Untracked:  code/primaryLift.out
    Untracked:  code/primaryLift.sh
    Untracked:  code/processhg38exons.py
    Untracked:  code/quantJunc.sh
    Untracked:  code/quantJunc_TEST.sh
    Untracked:  code/quantJunc_removeBad.sh
    Untracked:  code/quantLiftedPAS.err
    Untracked:  code/quantLiftedPAS.out
    Untracked:  code/quantLiftedPAS.sh
    Untracked:  code/quatJunc.err
    Untracked:  code/quatJunc.out
    Untracked:  code/recChimpback2Human.err
    Untracked:  code/recChimpback2Human.out
    Untracked:  code/recLiftchim2human.sh
    Untracked:  code/revLift.err
    Untracked:  code/revLift.out
    Untracked:  code/revLiftPAShg38to19.sh
    Untracked:  code/reverseLift.sh
    Untracked:  code/runCheckReverseLift.sh
    Untracked:  code/runChimpDiffIso.sh
    Untracked:  code/runChimpDiffIsoDF.sh
    Untracked:  code/runCountNucleotides.err
    Untracked:  code/runCountNucleotides.out
    Untracked:  code/runCountNucleotides.sh
    Untracked:  code/runCountNucleotidesPantro6.err
    Untracked:  code/runCountNucleotidesPantro6.out
    Untracked:  code/runCountNucleotides_pantro6.sh
    Untracked:  code/runFilterNumChroms.sh
    Untracked:  code/runHumanDiffIso.sh
    Untracked:  code/runHumanDiffIsoDF.sh
    Untracked:  code/runNuclearDiffIso_DF.sh
    Untracked:  code/runNuclearDifffIso.sh
    Untracked:  code/runTotalDiffIso.sh
    Untracked:  code/run_Chimpleafcutter_ds.err
    Untracked:  code/run_Chimpleafcutter_ds.out
    Untracked:  code/run_Chimpverifybam.err
    Untracked:  code/run_Chimpverifybam.out
    Untracked:  code/run_Humanleafcutter_dF.err
    Untracked:  code/run_Humanleafcutter_dF.out
    Untracked:  code/run_Humanleafcutter_ds.err
    Untracked:  code/run_Humanleafcutter_ds.out
    Untracked:  code/run_Nuclearleafcutter_ds.err
    Untracked:  code/run_Nuclearleafcutter_ds.out
    Untracked:  code/run_Nuclearleafcutter_dsDF.err
    Untracked:  code/run_Nuclearleafcutter_dsDF.out
    Untracked:  code/run_Totalleafcutter_ds.err
    Untracked:  code/run_Totalleafcutter_ds.out
    Untracked:  code/run_chimpverifybam.sh
    Untracked:  code/run_verifyBam.sh
    Untracked:  code/run_verifybam.err
    Untracked:  code/run_verifybam.out
    Untracked:  code/slurm-62824013.out
    Untracked:  code/slurm-62825841.out
    Untracked:  code/slurm-62826116.out
    Untracked:  code/slurm-64108209.out
    Untracked:  code/slurm-64108521.out
    Untracked:  code/slurm-64108557.out
    Untracked:  code/snakePASChimp.err
    Untracked:  code/snakePASChimp.out
    Untracked:  code/snakePAShuman.out
    Untracked:  code/snakemake.batch
    Untracked:  code/snakemakeChimp.err
    Untracked:  code/snakemakeChimp.out
    Untracked:  code/snakemakeHuman.err
    Untracked:  code/snakemakeHuman.out
    Untracked:  code/snakemakePAS.batch
    Untracked:  code/snakemakePASFiltChimp.err
    Untracked:  code/snakemakePASFiltChimp.out
    Untracked:  code/snakemakePASFiltHuman.err
    Untracked:  code/snakemakePASFiltHuman.out
    Untracked:  code/snakemakePASchimp.batch
    Untracked:  code/snakemakePAShuman.batch
    Untracked:  code/snakemake_chimp.batch
    Untracked:  code/snakemake_human.batch
    Untracked:  code/snakemakefiltPAS.batch
    Untracked:  code/snakemakefiltPAS_chimp.sh
    Untracked:  code/snakemakefiltPAS_human.sh
    Untracked:  code/spliceSite2Fasta.py
    Untracked:  code/submit-snakemake-chimp.sh
    Untracked:  code/submit-snakemake-human.sh
    Untracked:  code/submit-snakemakePAS-chimp.sh
    Untracked:  code/submit-snakemakePAS-human.sh
    Untracked:  code/submit-snakemakefiltPAS-chimp.sh
    Untracked:  code/submit-snakemakefiltPAS-human.sh
    Untracked:  code/subset_diffisopheno.py
    Untracked:  code/subset_diffisopheno_Chimp_tvN.py
    Untracked:  code/subset_diffisopheno_Chimp_tvN_DF.py
    Untracked:  code/subset_diffisopheno_Huma_tvN.py
    Untracked:  code/subset_diffisopheno_Huma_tvN_DF.py
    Untracked:  code/subset_diffisopheno_Nuclear_HvC.py
    Untracked:  code/subset_diffisopheno_Nuclear_HvC_DF.py
    Untracked:  code/subset_diffisopheno_Total_HvC.py
    Untracked:  code/test
    Untracked:  code/threeprimeOrthoFC.out
    Untracked:  code/threeprimeOrthoFC.sh
    Untracked:  code/threeprimeOrthoFCcd.err
    Untracked:  code/transcriptDTplotsNuclear.sh
    Untracked:  code/transcriptDTplotsTotal.sh
    Untracked:  code/verifyBam4973.sh
    Untracked:  code/verifyBam4973inHuman.sh
    Untracked:  code/verifybam4973.err
    Untracked:  code/verifybam4973.out
    Untracked:  code/verifybam4973HumanMap.err
    Untracked:  code/verifybam4973HumanMap.out
    Untracked:  code/wrap_Chimpverifybam.err
    Untracked:  code/wrap_Chimpverifybam.out
    Untracked:  code/wrap_chimpverifybam.sh
    Untracked:  code/wrap_verifyBam.sh
    Untracked:  code/wrap_verifybam.err
    Untracked:  code/wrap_verifybam.out
    Untracked:  code/writeMergecode.py
    Untracked:  data/._.DS_Store
    Untracked:  data/._HC_filenames.txt
    Untracked:  data/._HC_filenames.txt.sb-4426323c-IKIs0S
    Untracked:  data/._HC_filenames.xlsx
    Untracked:  data/._MapPantro6_meta.txt
    Untracked:  data/._MapPantro6_meta.txt.sb-a5794dd2-Cskmlm
    Untracked:  data/._MapPantro6_meta.xlsx
    Untracked:  data/._OppositeSpeciesMap.txt
    Untracked:  data/._OppositeSpeciesMap.txt.sb-a5794dd2-mayWJf
    Untracked:  data/._OppositeSpeciesMap.xlsx
    Untracked:  data/._RNASEQ_metadata.txt
    Untracked:  data/._RNASEQ_metadata.txt.sb-4426323c-TE4ns3
    Untracked:  data/._RNASEQ_metadata.txt.sb-51f67ae1-HXp7Gq
    Untracked:  data/._RNASEQ_metadata_2Removed.txt
    Untracked:  data/._RNASEQ_metadata_2Removed.txt.sb-4426323c-a4lBwx
    Untracked:  data/._RNASEQ_metadata_2Removed.xlsx
    Untracked:  data/._RNASEQ_metadata_stranded.txt
    Untracked:  data/._RNASEQ_metadata_stranded.txt.sb-a5794dd2-D659m2
    Untracked:  data/._RNASEQ_metadata_stranded.txt.sb-a5794dd2-ImNMoY
    Untracked:  data/._RNASEQ_metadata_stranded.txt.sb-e4bf31f0-ZGnGgl
    Untracked:  data/._RNASEQ_metadata_stranded.xlsx
    Untracked:  data/._metadata_HCpanel.txt
    Untracked:  data/._metadata_HCpanel.txt.sb-a3d92a2d-b9cYoF
    Untracked:  data/._metadata_HCpanel.txt.sb-a5794dd2-i594qs
    Untracked:  data/._metadata_HCpanel.txt.sb-f4823d1e-qihGek
    Untracked:  data/._metadata_HCpanel.xlsx
    Untracked:  data/._metadata_HCpanel_frompantro5.xlsx
    Untracked:  data/._~$RNASEQ_metadata.xlsx
    Untracked:  data/._~$metadata_HCpanel.xlsx
    Untracked:  data/._.xlsx
    Untracked:  data/BaseComp/
    Untracked:  data/CompapaQTLpas/
    Untracked:  data/DNDS/
    Untracked:  data/DTmatrix/
    Untracked:  data/DiffExpression/
    Untracked:  data/DiffIso_Nuclear/
    Untracked:  data/DiffIso_Nuclear_DF/
    Untracked:  data/DiffIso_Total/
    Untracked:  data/DiffSplice/
    Untracked:  data/DiffSplice_liftedJunc/
    Untracked:  data/DiffSplice_removeBad/
    Untracked:  data/DominantPAS/
    Untracked:  data/DominantPAS_DF/
    Untracked:  data/EvalPantro5/
    Untracked:  data/HC_filenames.txt
    Untracked:  data/HC_filenames.xlsx
    Untracked:  data/Khan_prot/
    Untracked:  data/Li_eqtls/
    Untracked:  data/MapPantro6_meta.txt
    Untracked:  data/MapPantro6_meta.xlsx
    Untracked:  data/MapStats/
    Untracked:  data/NormalizedClusters/
    Untracked:  data/NuclearHvC/
    Untracked:  data/NuclearHvC_DF/
    Untracked:  data/OppositeSpeciesMap.txt
    Untracked:  data/OppositeSpeciesMap.xlsx
    Untracked:  data/OverlapBenchmark/
    Untracked:  data/PAS/
    Untracked:  data/PAS_doubleFilter/
    Untracked:  data/Peaks_5perc/
    Untracked:  data/Pheno_5perc/
    Untracked:  data/Pheno_5perc_DF_nuclear/
    Untracked:  data/Pheno_5perc_nuclear/
    Untracked:  data/Pheno_5perc_nuclear_old/
    Untracked:  data/Pheno_5perc_total/
    Untracked:  data/PhyloP/
    Untracked:  data/RNASEQ_metadata.txt
    Untracked:  data/RNASEQ_metadata_2Removed.txt
    Untracked:  data/RNASEQ_metadata_2Removed.xlsx
    Untracked:  data/RNASEQ_metadata_stranded.txt
    Untracked:  data/RNASEQ_metadata_stranded.txt.sb-e4bf31f0-ZGnGgl/
    Untracked:  data/RNASEQ_metadata_stranded.xlsx
    Untracked:  data/SignalSites/
    Untracked:  data/SignalSites_doublefilter/
    Untracked:  data/SpliceSite/
    Untracked:  data/Threeprime2Ortho/
    Untracked:  data/TotalHvC/
    Untracked:  data/TwoBadSampleAnalysis/
    Untracked:  data/Wang_ribo/
    Untracked:  data/apaQTLGenes/
    Untracked:  data/bioGRID/
    Untracked:  data/chainFiles/
    Untracked:  data/cleanPeaks_anno/
    Untracked:  data/cleanPeaks_byspecies/
    Untracked:  data/cleanPeaks_lifted/
    Untracked:  data/files4viz_nuclear/
    Untracked:  data/files4viz_nuclear_DF/
    Untracked:  data/gviz/
    Untracked:  data/leafviz/
    Untracked:  data/liftover_files/
    Untracked:  data/mediation/
    Untracked:  data/mediation_DF/
    Untracked:  data/metadata_HCpanel.txt
    Untracked:  data/metadata_HCpanel.xlsx
    Untracked:  data/metadata_HCpanel_frompantro5.txt
    Untracked:  data/metadata_HCpanel_frompantro5.xlsx
    Untracked:  data/primaryLift/
    Untracked:  data/reverseLift/
    Untracked:  data/~$RNASEQ_metadata.xlsx
    Untracked:  data/~$metadata_HCpanel.xlsx
    Untracked:  data/.xlsx
    Untracked:  output/._.DS_Store
    Untracked:  output/dtPlots/
    Untracked:  projectNotes.Rmd
    Untracked:  proteinModelSet.Rmd

Unstaged changes:
    Modified:   analysis/DiffUsedIntronic.Rmd
    Modified:   analysis/ExploredAPA.Rmd
    Modified:   analysis/ExploredAPA_DF.Rmd
    Modified:   analysis/OppositeMap.Rmd
    Modified:   analysis/SpliceSiteStrength.Rmd
    Modified:   analysis/TotalVNuclearBothSpecies.Rmd
    Modified:   analysis/annotationInfo.Rmd
    Modified:   analysis/changeMisprimcut.Rmd
    Modified:   analysis/comp2apaQTLPAS.Rmd
    Modified:   analysis/correlationPhenos.Rmd
    Modified:   analysis/dAPA_Conservation.Rmd
    Modified:   analysis/dAPAandapaQTL_DF.Rmd
    Modified:   analysis/establishCutoffs.Rmd
    Modified:   analysis/investigatePantro5.Rmd
    Modified:   analysis/multiMap.Rmd
    Modified:   analysis/speciesSpecific.Rmd
    Modified:   analysis/speciesSpecific_DF.Rmd
    Modified:   analysis/upsetter_DF.Rmd

Note that any generated files, e.g. HTML, png, CSS, etc., are not included in this status report because it is ok for generated content to have uncommitted changes.

These are the previous versions of the R Markdown and HTML files. If you’ve configured a remote Git repository (see ?wflow_git_remote), click on the hyperlinks in the table below to view them.

File	Version	Author	Date	Message
Rmd	bec8ae9	brimittleman	2020-03-17	add mis filter annotation and pheno

library(tidyverse)

── Attaching packages ───────────────────────────────────────────────────────────────── tidyverse 1.2.1 ──

✔ ggplot2 3.1.1       ✔ purrr   0.3.2  
✔ tibble  2.1.1       ✔ dplyr   0.8.0.1
✔ tidyr   0.8.3       ✔ stringr 1.3.1  
✔ readr   1.3.1       ✔ forcats 0.3.0

── Conflicts ──────────────────────────────────────────────────────────────────── tidyverse_conflicts() ──
✖ dplyr::filter() masks stats::filter()
✖ dplyr::lag()    masks stats::lag()

library("gplots")


Attaching package: 'gplots'

The following object is masked from 'package:stats':

    lowess

library("R.utils")

Loading required package: R.oo

Loading required package: R.methodsS3

R.methodsS3 v1.7.1 (2016-02-15) successfully loaded. See ?R.methodsS3 for help.

R.oo v1.22.0 (2018-04-21) successfully loaded. See ?R.oo for help.


Attaching package: 'R.oo'

The following objects are masked from 'package:methods':

    getClasses, getMethods

The following objects are masked from 'package:base':

    attach, detach, gc, load, save

R.utils v2.7.0 successfully loaded. See ?R.utils for help.


Attaching package: 'R.utils'

The following object is masked from 'package:tidyr':

    extract

The following object is masked from 'package:utils':

    timestamp

The following objects are masked from 'package:base':

    cat, commandArgs, getOption, inherits, isOpen, parse, warnings

library("edgeR")

Loading required package: limma

library("limma")
library("scales")


Attaching package: 'scales'

The following object is masked from 'package:purrr':

    discard

The following object is masked from 'package:readr':

    col_factor

library("RColorBrewer")
library(reshape2)


Attaching package: 'reshape2'

The following object is masked from 'package:tidyr':

    smiths

Annotate and make phenotype

I am recreating the code from the annotation Like the PAS liftover, R code is from the compapa directory and bash code is done in the specific misprime directory.

mkdir ../data/cleanPeaks_anno
bedtools map -a ../data/cleanPeaks_lifted/AllPAS_postLift.sort.bed -b  /project2/gilad/briana/genome_anotation_data/hg38_refseq_anno/hg38_ncbiRefseq_Formatted_Allannotation.sort.bed -c 4 -S -o distinct > ../data/cleanPeaks_anno/AllPAS_postLift.sort_LocAnno.bed 

cp ../../Comparative_APA/code/chooseAnno2Bed.py .

python chooseAnno2Bed.py ../data/cleanPeaks_anno/AllPAS_postLift.sort_LocAnno.bed  ../data/cleanPeaks_anno/AllPAS_postLift.sort_LocAnnoPARSED.bed 

cp ../../Misprime4/code/LiftOrthoPAS2chimp.sh . #new dir  ../../Comparative_APA/data/chainFiles/

sbatch LiftOrthoPAS2chimp.sh

cp ../../Comparative_APA/code/bed2SAF_gen.py .

python bed2SAF_gen.py ../data/cleanPeaks_anno/AllPAS_postLift.sort_LocAnnoPARSED.bed  ../data/cleanPeaks_anno/AllPAS_postLift.sort_LocAnnoPARSED.SAF

python bed2SAF_gen.py ../data/cleanPeaks_anno/AllPAS_postLift.sort_LocAnnoPARSED_chimpLoc.bed  ../data/cleanPeaks_anno/AllPAS_postLift.sort_LocAnnoPARSED_chimpLoc.SAF


mkdir  ../Human/data/CleanLiftedPeaks_FC/
mkdir ../Chimp/data/CleanLiftedPeaks_FC/

cp ../../Comparative_APA/code/quantLiftedPAS.sh .


sbatch quantLiftedPAS.sh 


cp ../../Comparative_APA/code/fixFChead_bothfrac.py .


python fixFChead_bothfrac.py ../Human/data/CleanLiftedPeaks_FC/ALLPAS_postLift_LocParsed_Human ../Human/data/CleanLiftedPeaks_FC/ALLPAS_postLift_LocParsed_Human_fixed.fc


python fixFChead_bothfrac.py ../Chimp/data/CleanLiftedPeaks_FC/ALLPAS_postLift_LocParsed_Chimp ../Chimp/data/CleanLiftedPeaks_FC/ALLPAS_postLift_LocParsed_Chimp_fixed.fc

#make file ID
cp ../../Comparative_APA/code/makeFileID.py .

python makeFileID.py ../Chimp/data/CleanLiftedPeaks_FC/ALLPAS_postLift_LocParsed_Chimp ../Chimp/data/CleanLiftedPeaks_FC/ChimpFileID.txt

python makeFileID.py ../Human/data/CleanLiftedPeaks_FC/ALLPAS_postLift_LocParsed_Human ../Human/data/CleanLiftedPeaks_FC/HumanFileID.txt


mkdir ../Human/data/phenotype/
mkdir ../Chimp/data/phenotype/

cp ../../Comparative_APA/code/makePheno.py .

python makePheno.py  ../Human/data/CleanLiftedPeaks_FC/ALLPAS_postLift_LocParsed_Human_fixed.fc ../Human/data/CleanLiftedPeaks_FC/HumanFileID.txt ../Human/data/phenotype/ALLPAS_postLift_LocParsed_Human_Pheno.txt

python makePheno.py  ../Chimp/data/CleanLiftedPeaks_FC/ALLPAS_postLift_LocParsed_Chimp_fixed.fc ../Chimp/data/CleanLiftedPeaks_FC/ChimpFileID.txt ../Chimp/data/phenotype/ALLPAS_postLift_LocParsed_Chimp_Pheno.txt


cp ../../Comparative_APA/code/pheno2countonly.R .

Rscript pheno2countonly.R -I ../Human/data/phenotype/ALLPAS_postLift_LocParsed_Human_Pheno.txt -O ../Human/data/phenotype/ALLPAS_postLift_LocParsed_Human_Pheno_countOnly.txt

Rscript pheno2countonly.R -I ../Chimp/data/phenotype/ALLPAS_postLift_LocParsed_Chimp_Pheno.txt -O ../Chimp/data/phenotype/ALLPAS_postLift_LocParsed_Chimp_Pheno_countOnly.txt

cp ../../Comparative_APA/code/convertNumeric.py .

python convertNumeric.py ../Human/data/phenotype/ALLPAS_postLift_LocParsed_Human_Pheno_countOnly.txt ../Human/data/phenotype/ALLPAS_postLift_LocParsed_Human_Pheno_countOnlyNumeric.txt

python convertNumeric.py ../Chimp/data/phenotype/ALLPAS_postLift_LocParsed_Chimp_Pheno_countOnly.txt ../Chimp/data/phenotype/ALLPAS_postLift_LocParsed_Chimp_Pheno_countOnlyNumeric.txt

Expression cutoff.

I will use the same cutoff as I used in the original data.

humanPAS=read.table("../../Misprime6/Human/data/CleanLiftedPeaks_FC/ALLPAS_postLift_LocParsed_Human_fixed.fc", header=T, stringsAsFactors = F) %>% 
  separate(Geneid, into=c("disc","PAS","chrom", "start","end","strand","geneid"), sep=":") %>%
  separate(geneid,into=c("gene","loc"),sep="_") %>%
  dplyr::select(gene,contains("_N")) %>%
  gather(key="ind", value="count", -gene) %>% 
  group_by(ind, gene) %>%
  summarize(GeneCount=sum(count)) %>% 
  spread(ind, GeneCount)

Warning: Expected 2 pieces. Additional pieces discarded in 2 rows [27653,
52819].

chimpPAS=read.table("../../Misprime6/Chimp/data/CleanLiftedPeaks_FC/ALLPAS_postLift_LocParsed_Chimp_fixed.fc", header=T, stringsAsFactors = F) %>% 
  separate(Geneid, into=c("disc","PAS","chrom", "start","end","strand","geneid"), sep=":") %>%
  separate(geneid,into=c("gene","loc"),sep="_") %>%
  dplyr::select(gene,contains("_N")) %>%
  gather(key="ind", value="count", -gene) %>% 
  group_by(ind, gene) %>%
  summarize(GeneCount=sum(count)) %>% 
  spread(ind, GeneCount)

Warning: Expected 2 pieces. Additional pieces discarded in 2 rows [27653,
52819].

#can use the same meta becuase it is ordered the same  
metadata=read.table("../data/metadata_HCpanel.txt",header = T) %>% mutate(id2=ifelse(grepl("pt", ID), ID, paste("X", ID, sep=""))) %>% filter(Fraction=="Nuclear")

order=c(metadata$id2[1:10], "pt30_N", "pt91_N")

BothbyGene= chimpPAS %>% inner_join(humanPAS,by="gene") %>% dplyr::select(gene,order)

#count matrix:
Genematrix=as.matrix(BothbyGene %>% column_to_rownames(var="gene"))

colors <- colorRampPalette(c(brewer.pal(9, "Blues")[1],brewer.pal(9, "Blues")[9]))(100)

pal <- c(brewer.pal(9, "Set1"), brewer.pal(8, "Set2"), brewer.pal(12, "Set3"))
labels <- paste(metadata$Species,metadata$Line, sep=" ")

# Clustering (original code from Julien Roux)
cors <- cor(Genematrix, method="spearman", use="pairwise.complete.obs")


heatmap.2( cors, scale="none", col = colors, margins = c(12, 12), trace='none', denscol="white", labCol=labels, ColSideColors=pal[as.integer(as.factor(metadata$Species))], RowSideColors=pal[as.integer(as.factor(metadata$Collection))+9], cexCol = 0.2 + 1/log10(15), cexRow = 0.2 + 1/log10(15))

log_counts_genes <- as.data.frame(log2(Genematrix))
plotDensities(log_counts_genes, col=pal[as.numeric(metadata$Species)], legend="topright")

cpm <- cpm(Genematrix, log=TRUE)
plotDensities(cpm, col=pal[as.numeric(metadata$Species)], legend="topright")

## Create edgeR object (dge) to calculate TMM normalization  
dge_original <- DGEList(counts=as.matrix(Genematrix), genes=rownames(Genematrix), group = as.character(t(labels)))
dge_original <- calcNormFactors(dge_original)

tmm_cpm <- cpm(dge_original, normalized.lib.sizes=TRUE, log=TRUE, prior.count = 0.25)
head(cpm)

         X18498_N   X18499_N   X18502_N X18504_N    X18510_N   X18523_N
A1BG     5.983536  5.4183778  5.4098495 4.663320  4.48324922 5.62423862
A1BG-AS1 4.071747  3.4978452  4.1234882 3.262501  3.72249613 3.57914708
A2M      2.142925 -1.5743299 -0.4993838 2.312121 -0.03058551 0.01319229
A4GALT   5.543397 -0.5366276  2.9226146 3.063796  3.91933502 4.74234267
AAAS     4.417342  3.6273229  4.6991559 5.099780  4.44115086 3.65822441
AACS     6.288806  7.4218057  5.7257025 7.208370  6.62098854 6.61684685
         X18358_N    X3622_N  X3659_N    X4973_N     pt30_N     pt91_N
A1BG     6.457101  4.3145111 5.870401  3.4115203  5.5275684  4.9061884
A1BG-AS1 4.355529  2.0046092 3.869027  0.9238313  1.8067853  1.2271781
A2M      6.023206  6.9373362 7.220784  5.6298885  5.4570591  5.0627709
A4GALT   3.710202 -0.1138492 3.898710 -0.3143677 -0.1748582 -0.1380633
AAAS     4.039505  4.4757803 4.758339  4.2295650  4.5165243  4.1312581
AACS     6.002210  5.3341952 5.224647  5.4353906  5.3247138  5.2857423

log2cpm plot

plotDensities(tmm_cpm, col=pal[as.numeric(metadata$Species)], legend="topright")

keep.exprs=rowSums(tmm_cpm>2) >8

counts_filtered= Genematrix[keep.exprs,]




plotDensities(counts_filtered, col=pal[as.numeric(metadata$Species)], legend="topright")

labels <- paste(metadata$Species, metadata$Line, sep=" ")
dge_in_cutoff <- DGEList(counts=as.matrix(counts_filtered), genes=rownames(counts_filtered), group = as.character(t(labels)))
dge_in_cutoff <- calcNormFactors(dge_in_cutoff)

cpm_in_cutoff <- cpm(dge_in_cutoff, normalized.lib.sizes=TRUE, log=TRUE, prior.count = 0.25)
head(cpm_in_cutoff)

         X18498_N X18499_N X18502_N X18504_N X18510_N X18523_N X18358_N
A1BG     5.957948 5.443869 5.490546 4.616405 4.354045 5.783591 6.492871
A1BG-AS1 4.027659 3.495425 4.189664 3.187911 3.580187 3.711004 4.375357
AAAS     4.378646 3.628186 4.773447 5.057221 4.311390 3.792034 4.054206
AACS     6.264487 7.454726 5.808372 7.175235 6.506229 6.780525 6.036193
AAGAB    6.732123 5.570277 5.833618 6.075778 5.897792 5.243105 6.296532
AAK1     7.239137 7.075132 7.262894 6.601016 6.875455 7.462096 7.348393
          X3622_N  X3659_N   X4973_N   pt30_N   pt91_N
A1BG     4.517128 5.929125 3.5521872 5.756699 5.035251
A1BG-AS1 2.117826 3.905766 0.8602056 1.916882 1.175998
AAAS     4.680711 4.808601 4.3880096 4.736102 4.250081
AACS     5.547867 5.279265 5.6068936 5.552428 5.418120
AAGAB    6.048714 6.023814 5.8432555 5.820213 5.804474
AAK1     6.456939 6.907735 7.8154504 6.295812 6.384961

GenesCutoff=rownames(cpm_in_cutoff)
NormalizedGenesCuttoff=as.data.frame(cbind(Gene_stable_ID=GenesCutoff, cpm_in_cutoff))

hist(cpm_in_cutoff, xlab = "Log2(CPM)", main = "Log2(CPM) values for genes meeting the filtering criteria", breaks = 100 )

Species <- factor(metadata$Species)
design <- model.matrix(~ 0 + Species)
head(design)

  SpeciesChimp SpeciesHuman
1            0            1
2            0            1
3            0            1
4            0            1
5            0            1
6            0            1

colnames(design) <- gsub("Species", "", dput(colnames(design)))

c("SpeciesChimp", "SpeciesHuman")

cpm.voom<- voom(counts_filtered, design, normalize.method="quantile", plot=T)

boxplot(cpm.voom$E, col = pal[as.numeric(metadata$Species)],las=2)

plotDensities(cpm.voom, col =  pal[as.numeric(metadata$Species)], legend = "topleft")

length(GenesCutoff)

[1] 9428

GenesCutoffDF=as.data.frame(GenesCutoff) %>% rename("genes"=GenesCutoff)
#mkdir ../data/OverlapBenchmark
write.table(GenesCutoffDF,"../../Misprime6/data/OverlapBenchmark/genesPassingCuttoff.txt", col.names = T, row.names = F,quote = F)

Filter PAS on these genes and 5%

HumanAnno=read.table("../../Misprime6/Human/data/phenotype/ALLPAS_postLift_LocParsed_Human_Pheno.txt", header = T, stringsAsFactors = F) %>% tidyr::separate(chrom, sep = ":", into = c("chr", "start", "end", "id")) %>% tidyr::separate(id, sep="_", into=c("gene", "strand", "peak"))  %>% separate(peak,into=c("loc", "disc","PAS"), sep="-")
IndH=colnames(HumanAnno)[9:ncol(HumanAnno)]

HumanUsage=read.table("../../Misprime6/Human/data/phenotype/ALLPAS_postLift_LocParsed_Human_Pheno_countOnlyNumeric.txt", col.names = IndH)

HumanMean=as.data.frame(cbind(HumanAnno[,1:8], Human=rowMeans(HumanUsage)))

HumanUsage_anno=as.data.frame(cbind(HumanAnno[,1:8],HumanUsage ))

ChimpAnno=read.table("../../Misprime6/Chimp/data/phenotype/ALLPAS_postLift_LocParsed_Chimp_Pheno.txt", header = T, stringsAsFactors = F) %>% tidyr::separate(chrom, sep = ":", into = c("chr", "start", "end", "id")) %>% tidyr::separate(id, sep="_", into=c("gene", "strand", "peak"))  %>% separate(peak,into=c("loc", "disc","PAS"), sep="-")
IndC=colnames(ChimpAnno)[9:ncol(ChimpAnno)]

ChimpUsage=read.table("../../Misprime6/Chimp/data/phenotype/ALLPAS_postLift_LocParsed_Chimp_Pheno_countOnlyNumeric.txt", col.names = IndC)

ChimpMean=as.data.frame(cbind(ChimpAnno[,1:8], Chimp=rowMeans(ChimpUsage)))

ChimpUsage_anno=as.data.frame(cbind(ChimpAnno[,1:8],ChimpUsage ))

BothMean=ChimpMean %>% full_join(HumanMean, by=c("chr","start","end","gene"   ,"strand", "loc", "disc","PAS" )) 

BothMeanM=melt(BothMean,id.vars =c("chr","start","end","gene"   ,"strand", "loc", "disc","PAS" ),variable.name = "Species", value.name = "MeanUsage" ) %>% filter(loc !="008559", loc != "009911")

ggplot(BothMeanM, aes(x=loc, y=MeanUsage,by=Species,fill=Species)) + geom_boxplot()  + scale_fill_brewer(palette = "Dark2")

ggplot(BothMeanM, aes(x=MeanUsage,by=Species,col=Species)) + stat_ecdf(geom = "point", alpha=.25)  + scale_color_brewer(palette = "Dark2") + labs(title="Cumulative Distribution plot for PAS Usage", x="Mean Usage- both fractions", y="F(Mean Usage)")

Implement cutoffs for gene expression and usage.

BothMean_5= BothMean %>% filter(Chimp >=0.05 | Human >= 0.05,gene %in% GenesCutoffDF$genes)

BothMean_5M=melt(BothMean_5,id.vars =c("chr","start","end","gene"   ,"strand", "loc", "disc","PAS" ),variable.name = "Species", value.name = "MeanUsage" ) %>% filter(loc !="008559")

ggplot(BothMean_5M, aes(x=loc, y=MeanUsage,by=Species,fill=Species)) + geom_boxplot()  + scale_fill_brewer(palette = "Dark2")

ggplot(BothMean_5M, aes(x=MeanUsage,by=Species,col=Species)) + stat_ecdf(geom = "point", alpha=.25)  + scale_color_brewer(palette = "Dark2") + labs(title="Cumulative Distribution plot for PAS Usage at 5%", x="Mean Usage- both fractions", y="F(Mean Usage)")

ggplot(BothMean_5M, aes(x=MeanUsage,by=Species,fill=Species)) + geom_histogram(alpha=.5, bins=30, position = "dodge")  + scale_fill_brewer(palette = "Dark2")

mkdir ../data/Peaks_5perc
mkdir ../data/Pheno_5perc

BothMean_5_out=BothMean_5 %>% dplyr::select(PAS,disc, gene, loc,chr, start, end,Chimp, Human)
write.table(BothMean_5_out, "../../Misprime6/data/Peaks_5perc/Peaks_5perc_either_bothUsage.txt", row.names = F, col.names = T, quote = F)

BothMean_5_out_noUN=BothMean_5 %>% dplyr::select(PAS,disc, gene, loc,chr, start, end,Chimp, Human) %>% filter(!grepl("Un",chr))

write.table(BothMean_5_out_noUN, "../../Misprime6/data/Peaks_5perc/Peaks_5perc_either_bothUsage_noUnchr.txt", row.names = F, col.names = T, quote = F)
#write bed with human coord for igv
BothMean_5_bed=BothMean_5 %>% dplyr::select(chr, start, end, PAS, Human, strand)
write.table(BothMean_5_bed,"../../Misprime6/data/Peaks_5perc/Peaks_5perc_either_HumanCoordHummanUsage.bed", row.names = F, col.names = T, quote = F)

ggplot(BothMean_5_out, aes(x=disc, fill=disc))+  geom_bar(aes(y = (..count..)/sum(..count..)))+ scale_fill_brewer(palette = "Dark2")

BothMean_5_outmean= BothMean_5_out %>% mutate(meanUsage=(Human+Chimp)/2)
ggplot(BothMean_5M, aes(x=disc, by= Species, fill=Species, y=MeanUsage)) + geom_boxplot() + scale_y_log10()+ scale_fill_brewer(palette = "Dark2")

Warning: Transformation introduced infinite values in continuous y-axis

Warning: Removed 516 rows containing non-finite values (stat_boxplot).

ChimpUsage_anno_5perc= ChimpUsage_anno %>% filter(PAS %in% BothMean_5$PAS)

write.table(ChimpUsage_anno_5perc, "../../Misprime6/data/Pheno_5perc/Chimp_Pheno_5perc.txt", row.names = F, col.names = T, quote = F)

HumaUsage_anno_5perc= HumanUsage_anno %>% filter(PAS %in% BothMean_5$PAS)

write.table(HumaUsage_anno_5perc, "../../Misprime6/data/Pheno_5perc/Human_Pheno_5perc.txt", row.names = F, col.names = T, quote = F)

sessionInfo()

R version 3.5.1 (2018-07-02)
Platform: x86_64-pc-linux-gnu (64-bit)
Running under: Scientific Linux 7.4 (Nitrogen)

Matrix products: default
BLAS/LAPACK: /software/openblas-0.2.19-el7-x86_64/lib/libopenblas_haswellp-r0.2.19.so

locale:
 [1] LC_CTYPE=en_US.UTF-8       LC_NUMERIC=C              
 [3] LC_TIME=en_US.UTF-8        LC_COLLATE=en_US.UTF-8    
 [5] LC_MONETARY=en_US.UTF-8    LC_MESSAGES=en_US.UTF-8   
 [7] LC_PAPER=en_US.UTF-8       LC_NAME=C                 
 [9] LC_ADDRESS=C               LC_TELEPHONE=C            
[11] LC_MEASUREMENT=en_US.UTF-8 LC_IDENTIFICATION=C       

attached base packages:
[1] stats     graphics  grDevices utils     datasets  methods   base     

other attached packages:
 [1] reshape2_1.4.3     RColorBrewer_1.1-2 scales_1.0.0      
 [4] edgeR_3.24.0       limma_3.38.2       R.utils_2.7.0     
 [7] R.oo_1.22.0        R.methodsS3_1.7.1  gplots_3.0.1      
[10] forcats_0.3.0      stringr_1.3.1      dplyr_0.8.0.1     
[13] purrr_0.3.2        readr_1.3.1        tidyr_0.8.3       
[16] tibble_2.1.1       ggplot2_3.1.1      tidyverse_1.2.1   

loaded via a namespace (and not attached):
 [1] Rcpp_1.0.2         locfit_1.5-9.1     lubridate_1.7.4   
 [4] lattice_0.20-38    gtools_3.8.1       assertthat_0.2.0  
 [7] rprojroot_1.3-2    digest_0.6.18      R6_2.3.0          
[10] cellranger_1.1.0   plyr_1.8.4         backports_1.1.2   
[13] evaluate_0.12      httr_1.3.1         pillar_1.3.1      
[16] rlang_0.4.0        lazyeval_0.2.1     readxl_1.1.0      
[19] rstudioapi_0.10    gdata_2.18.0       whisker_0.3-2     
[22] rmarkdown_1.10     labeling_0.3       munsell_0.5.0     
[25] broom_0.5.1        compiler_3.5.1     httpuv_1.4.5      
[28] modelr_0.1.2       pkgconfig_2.0.2    htmltools_0.3.6   
[31] tidyselect_0.2.5   workflowr_1.6.0    crayon_1.3.4      
[34] withr_2.1.2        later_0.7.5        bitops_1.0-6      
[37] grid_3.5.1         nlme_3.1-137       jsonlite_1.6      
[40] gtable_0.2.0       git2r_0.26.1       magrittr_1.5      
[43] KernSmooth_2.23-15 cli_1.1.0          stringi_1.2.4     
[46] fs_1.3.1           promises_1.0.1     xml2_1.2.0        
[49] generics_0.0.2     tools_3.5.1        glue_1.3.0        
[52] hms_0.4.2          yaml_2.2.0         colorspace_1.3-2  
[55] caTools_1.17.1.1   rvest_0.3.2        knitr_1.20        
[58] haven_1.1.2

Annotate Misprime 6

Briana Mittleman

3/17/2020

Annotate and make phenotype

Expression cutoff.

Filter PAS on these genes and 5%