06d7be056190c14b85e71bc12523f18ea6815b5e markd Mon Dec 7 00:50:29 2020 -0800 BLAT mmap index support merge with master diff --git src/hg/utils/otto/sarscov2phylo/processRelease.sh src/hg/utils/otto/sarscov2phylo/processRelease.sh new file mode 100755 index 0000000..979fcc4 --- /dev/null +++ src/hg/utils/otto/sarscov2phylo/processRelease.sh @@ -0,0 +1,148 @@ +#!/bin/bash +set -beEu -x -o pipefail + +# Do not modify this script, modify the source tree copy: +# kent/src/hg/utils/otto/sarscov2phylo/processRelease.sh + +usage() { + echo "usage: $0 releaseLabel problematic_sites_sarsCov2.vcf" +} + +if [ $# != 2 ]; then + usage + exit 1 +fi + +releaseLabel=$1 +problematicSitesVcf=$2 + +echo "releaseLabel=$releaseLabel problematicSitesVcf=$problematicSitesVcf" + +scriptDir=$(dirname "${BASH_SOURCE[0]}") + +# UShER (and matToVcf) +usherDir=~angie/github/usher +usher=$usherDir/build/usher +matToVcf=$usherDir/build/matToVcf + +# strain_phylogenetics / find_parsimonious_assignments for parsimony scores +find_parsimonious_assignments=~angie/github/strain_phylogenetics/build/find_parsimonious_assignments + +# Files generated by getRelease.sh +metadata=gisaid_metadata.tsv +fasta=gisaid_sequences.fasta +alignedFasta=gisaid_aligned.fasta + +# We'll need to prune the tree to keep only those sequences that are in metadata and fasta: +grep ^\> $fasta | sed -re 's/^>//' | sort > namesInFasta + +# Make renaming files for tree (EPI_ IDs) and VCF (isolate names): add strain name and +# date with shortened year. +awk -F"\t" '{print $3 "\t" $1 "|" $3 "|" $5;}' $metadata \ +| grep -Fwf namesInFasta \ +| sed -re 's/\|20/\|/;' \ +| sort > tree.renaming +awk -F"\t" '{print $1 "\t" $1 "|" $3 "|" $5;}' $metadata \ +| grep -Fwf namesInFasta \ +| sed -re 's/\|20/\|/;' \ +| sort > vcf.renaming +wc -l tree.renaming +wc -l vcf.renaming + +# Rename tree IDs from just EPI IDs to strain|epi|date ids and prune any sequences not in +# metadata and fasta: +phyloRenameAndPrune ft_SH.tree tree.renaming ft_SH.renamed.tree +sed -re 's/\)[0-9.]+:/\):/g; s/:[0-9e.:-]+//g; s/[\(\);]//g; s/,/\n/g;'" s/'//g;" \ + ft_SH.renamed.tree \ +| sort > tree.renamedPruned.ids.sorted +wc -l tree.renamedPruned.ids.sorted + +# Set the root of the tree to the Wuhan/Hu-1 (NC_045512.2) reference not WIV04: +~angie/github/newick_utils/src/nw_reroot ft_SH.renamed.tree \ + 'Wuhan/Hu-1/2019|EPI_ISL_402125|19-12-26' > ft_SH.reroot.nh + +# Run faToVcf without any filtering, masking or conversion of ambiguous alleles: +time faToVcf $alignedFasta stdout -includeRef \ + -ref=Wuhan/Hu-1/2019 \ + -vcfChrom=NC_045512v2 -verbose=2 \ +| vcfRenameAndPrune stdin vcf.renaming gisaid-$releaseLabel.unfiltered.vcf +ls -l gisaid-$releaseLabel.unfiltered.vcf +wc -l gisaid-$releaseLabel.unfiltered.vcf +gzip -f gisaid-$releaseLabel.unfiltered.vcf + +# Run usher and matToVcf to make ambig-resolved VCF for hgTracks +time $usher -c \ + --vcf gisaid-$releaseLabel.unfiltered.vcf.gz \ + --tree ft_SH.reroot.nh \ + --save-mutation-annotated-tree sarscov2phylo-$releaseLabel.notMasked.pb \ + --write-uncondensed-final-tree +mv uncondensed-final-tree.nh ft_SH.reroot.collapsed.notMasked.nh +$matToVcf -i sarscov2phylo-$releaseLabel.notMasked.pb \ + -v gisaid-$releaseLabel.vcf +bgzip -f gisaid-$releaseLabel.vcf +tabix -p vcf gisaid-$releaseLabel.vcf.gz + +# Remove problematic sites recommended for masking for usher/phyloPlace +tawk '{ if ($1 ~ /^#/) { print; } else if ($7 == "mask") { $1 = "NC_045512v2"; print; } }' \ + $problematicSitesVcf > mask.vcf +time vcfFilter -excludeVcf=mask.vcf gisaid-$releaseLabel.unfiltered.vcf.gz \ +| gzip -c \ + > gisaid-$releaseLabel.unfiltered.masked.vcf.gz + +# Full find_parsimonious_assignments output on uncollapsed tree for collaborators +time $find_parsimonious_assignments --tree ft_SH.reroot.nh \ + --vcf gisaid-$releaseLabel.unfiltered.masked.vcf.gz \ +| gzip -c \ + > find_parsimonious_assignments.$releaseLabel.out.gz + +# Run usher to collapse tree and make masked protobuf for hgPhyloPlace +time $usher -c \ + --vcf gisaid-$releaseLabel.unfiltered.masked.vcf.gz \ + --tree ft_SH.reroot.nh \ + --save-mutation-annotated-tree sarscov2phylo-$releaseLabel.masked.pb \ + --write-uncondensed-final-tree +mv uncondensed-final-tree.nh ft_SH.reroot.collapsed.nh + +# Parsimony scores on collapsed tree +time $find_parsimonious_assignments --tree ft_SH.reroot.collapsed.nh \ + --vcf <(gunzip -c gisaid-$releaseLabel.vcf.gz) \ +| tail -n+2 \ +| sed -re 's/^[A-Z]([0-9]+)[A-Z,]+.*parsimony_score=([0-9]+).*/\1\t\2/;' \ +| tawk '{print "NC_045512v2", $1-1, $1, $2;}' \ +| sort -k2n,2n \ + > gisaid-$releaseLabel.parsimony.bg +bedGraphToBigWig gisaid-$releaseLabel.parsimony.bg /hive/data/genomes/wuhCor1/chrom.sizes \ + gisaid-$releaseLabel.parsimony.bw + +# Full find_parsimonious_assignments output on collapsed tree for collaborators +time $find_parsimonious_assignments --tree ft_SH.reroot.collapsed.nh \ + --vcf gisaid-$releaseLabel.unfiltered.masked.vcf.gz \ + > find_parsimonious_assignments.$releaseLabel.collapsed.out + +# Now filter by frequency for faster track display: alt al freq >= 0.001 +sampleCount=$(wc -l < tree.renamedPruned.ids.sorted) +minAc001=$(( (($sampleCount + 999) / 1000) )) +time vcfFilter -minAc=$minAc001 -rename gisaid-$releaseLabel.vcf.gz \ + > gisaid-$releaseLabel.minAf.001.vcf +ls -l gisaid-$releaseLabel.minAf.001.vcf +wc -l gisaid-$releaseLabel.minAf.001.vcf +bgzip -f gisaid-$releaseLabel.minAf.001.vcf +tabix -p vcf gisaid-$releaseLabel.minAf.001.vcf.gz + +# Alt al freq >= 0.01 +minAc01=$(( (($sampleCount + 99) / 100) )) +time vcfFilter -minAc=$minAc01 -rename gisaid-$releaseLabel.minAf.001.vcf.gz \ + > gisaid-$releaseLabel.minAf.01.vcf +ls -l gisaid-$releaseLabel.minAf.01.vcf +wc -l gisaid-$releaseLabel.minAf.01.vcf +bgzip -f gisaid-$releaseLabel.minAf.01.vcf +tabix -p vcf gisaid-$releaseLabel.minAf.01.vcf.gz + +# Make sample color files +#*** TODO: Nextstrain Clade is now in field 18! Add that as an option! +cut -f3,18-20 $metadata \ +| sort > metadata.epiToLineageAndClades +# Join on EPI ID to associate tree sample names with lineages. +join -t$'\t' tree.renaming metadata.epiToLineageAndClades -o 1.2,2.2,2.3,2.4 > sampleToLineage +$scriptDir/cladeLineageColors.pl sampleToLineage +gzip -f *Colors