############################################################################# ## 5-Way Multiz (DONE - 2016-12-17 - Hiram) ssh hgwdev mkdir /hive/data/genomes/hg38/bed/multiz5way cd /hive/data/genomes/hg38/bed/multiz5way # from the 218-way in the source tree, select out the 5 used here: /cluster/bin/phast/tree_doctor \ --prune-all-but hg38,galVar1,cavPor3,mm10,tupChi1 \ /cluster/home/hiram/kent/src/hg/utils/phyloTrees/218way.nh \ > hg38.5way.nh cat hg38.5way.nh # ((hg38:0.143908,(tupChi1:0.120000,galVar1:0.080000):0.054937):0.002000, # (mm10:0.315424,cavPor3:0.175779):0.041059); # no need for rearrangement since hg38 is already at the top # what that looks like: ~/kent/src/hg/utils/phyloTrees/asciiTree.pl hg38.5way.nh | sed -e 's/^/# /;' # ((hg38:0.143908, # (tupChi1:0.12, # galVar1:0.080000):0.054937):0.002000, # (mm10:0.315424, # cavPor3:0.175779):0.041059); # extract species list from that .nh file sed 's/[a-z][a-z]*_//g; s/:[0-9\.][0-9\.]*//g; s/;//; /^ *$/d' \ hg38.5way.nh | xargs echo | sed 's/ //g; s/,/ /g' \ | sed 's/[()]//g; s/,/ /g' | tr '[ ]' '[\n]' > species.list.txt # construct db to name translation list: cat species.list.txt | while read DB do hgsql -N -e "select name,organism from dbDb where name=\"${DB}\";" hgcentraltest done | sed -e "s/\t/->/; s/ /_/g;" | sed -e 's/$/;/' | sed -e 's/\./_/g' \ | sed -e 's/-nosed/_nosed/; s/-eating/_eating/;' > db.to.name.txt # construct a common name .nh file: /cluster/bin/phast/tree_doctor --rename \ "`cat db.to.name.txt`" hg38.5way.nh | sed -e 's/00*)/)/g; s/00*,/,/g' \ | $HOME/kent/src/hg/utils/phyloTrees/asciiTree.pl /dev/stdin \ > hg38.5way.commonNames.nh cat hg38.5way.commonNames.nh | sed -e 's/^/# /;' # ((Human:0.143908, # (Chinese_tree_shrew:0.12, # Malayan_flying_lemur:0.08):0.054937):0.002, # (Mouse:0.315424, # Guinea_pig:0.175779):0.041059); # Use this specification in the phyloGif tool: # http://genome.ucsc.edu/cgi-bin/phyloGif # to obtain a png image for src/hg/htdocs/images/phylo/hg38_5way.png ~/kent/src/hg/utils/phyloTrees/asciiTree.pl hg38.5way.nh > t.nh ~/kent/src/hg/utils/phyloTrees/scientificNames.sh t.nh \ | $HOME/kent/src/hg/utils/phyloTrees/asciiTree.pl /dev/stdin \ > hg38.5way.scientificNames.nh rm -f t.nh cat hg38.5way.scientificNames.nh | sed -e 's/^/# /;' # ((Homo_sapiens:0.143908, # (Tupaia_chinensis:0.12, # Galeopterus_variegatus:0.08):0.054937):0.002, # (Mus_musculus:0.315424, # Cavia_porcellus:0.175779):0.041059); /cluster/bin/phast/all_dists hg38.5way.nh | grep hg38 \ | sed -e "s/hg38.//" | sort -k2n > 5way.distances.txt # Use this output to create the table below cat 5way.distances.txt | sed -e 's/^/# /;' # galVar1 0.278845 # tupChi1 0.318845 # cavPor3 0.362746 # mm10 0.502391 printf '#!/usr/bin/env perl use strict; use warnings; open (FH, "<5way.distances.txt") or die "can not read 5way.distances.txt"; my $count = 0; while (my $line = ) { chomp $line; my ($D, $dist) = split('"'"'\\s+'"'"', $line); my $chain = "chain" . ucfirst($D); my $B="/hive/data/genomes/hg38/bed/lastz.$D/fb.hg38." . $chain . "Link.txt"; my $chainLinkMeasure = `awk '"'"'{print \\$5}'"'"' ${B} 2> /dev/null | sed -e "s/(//; s/)//"`; chomp $chainLinkMeasure; $chainLinkMeasure = 0.0 if (length($chainLinkMeasure) < 1); $chainLinkMeasure =~ s/\\%%//; my $swapFile="/hive/data/genomes/${D}/bed/lastz.hg38/fb.${D}.chainHg38Link.txt"; my $swapMeasure = "N/A"; if ( -s $swapFile ) { $swapMeasure = `awk '"'"'{print \\$5}'"'"' ${swapFile} 2> /dev/null | sed -e "s/(//; s/)//"`; chomp $swapMeasure; $swapMeasure = 0.0 if (length($swapMeasure) < 1); $swapMeasure =~ s/\\%%//; } my $orgName= `hgsql -N -e "select organism from dbDb where name='"'\$D'"';" hgcentraltest`; chomp $orgName; if (length($orgName) < 1) { $orgName="N/A"; } ++$count; printf "# %%02d %%.4f (%%%% %%05.3f) (%%%% %%05.3f) - %%s %%s\\n", $count, $dist, $chainLinkMeasure, $swapMeasure, $orgName, $D; } close (FH); ' > sizeStats.pl chmod +x ./sizeStats.pl ./sizeStats.pl # If you can fill in all the numbers in this table, you are ready for # the multiple alignment procedure # featureBits chainLink measures # chainLink # N distance on hg38 on other other species # 01 0.2788 (% 57.036) (% 64.881) - Malayan flying lemur galVar1 # 02 0.3188 (% 45.256) (% 50.350) - Chinese tree shrew tupChi1 # 03 0.3627 (% 42.371) (% 48.000) - Guinea pig cavPor3 # 04 0.5024 (% 31.653) (% 35.372) - Mouse mm10 # None of this concern for distances matters in building the first step, the # maf files. The distances will be better calibrated later. # create species list and stripped down tree for autoMZ sed 's/[a-z][a-z]*_//g; s/:[0-9\.][0-9\.]*//g; s/;//; /^ *$/d' \ hg38.5way.nh | xargs echo | sed 's/ //g; s/,/ /g' > tree.nh sed 's/[()]//g; s/,/ /g' tree.nh > species.list # hg38 tupChi1 galVar1 mm10 cavPor3 # survey N50 for each for db in `cat species.list` do n50.pl /hive/data/genomes/$db/chrom.sizes done # reading: /hive/data/genomes/hg38/chrom.sizes # contig count: 455, total size: 3209286105, one half size: 1604643052 # cumulative N50 count contig contig size 1547391171 8 chrX 156040895 1604643052 one half size 1692529807 9 chr8 145138636 # reading: /hive/data/genomes/tupChi1/chrom.sizes # contig count: 50750, total size: 2846580235, one half size: 1423290117 # cumulative N50 count contig contig size 1419920836 231 KB321095 3691413 1423290117 one half size 1423590960 232 KB321106 3670124 # reading: /hive/data/genomes/galVar1/chrom.sizes # contig count: 179514, total size: 3187660572, one half size: 1593830286 # cumulative N50 count contig contig size 1593691350 3422 NW_007730159v1 245222 1593830286 one half size 1593936539 3423 NW_007729331v1 245189 # reading: /hive/data/genomes/mm10/chrom.sizes # contig count: 66, total size: 2730871774, one half size: 1365435887 # cumulative N50 count contig contig size 1312176979 8 chr7 145441459 1365435887 one half size 1442871972 9 chr10 130694993 # reading: /hive/data/genomes/cavPor3/chrom.sizes # contig count: 3144, total size: 2723219641, one half size: 1361609820 # cumulative N50 count contig contig size 1356838683 27 scaffold_25 28222655 1361609820 one half size 1384780737 28 scaffold_27 27942054 # bash shell syntax here ... cd /hive/data/genomes/hg38/bed/multiz5way export H=/hive/data/genomes/hg38/bed mkdir mafLinks # good assemblies can use syntenic net: # galVar1 mm10 cavPor3 tupChi1 for G in galVar1 mm10 cavPor3 tupChi1 do mkdir mafLinks/$G echo ln -s ${H}/lastz.$G/axtChain/hg38.${G}.synNet.maf.gz ./mafLinks/$G ln -s ${H}/lastz.$G/axtChain/hg38.${G}.synNet.maf.gz ./mafLinks/$G done # other assemblies using recip best net: # verify the symLinks are good: ls -ogrtL mafLinks/*/* | sed -e 's/^/# /; s/-rw-rw-r-- 1//;' # 999907313 Feb 19 2015 mafLinks/tupChi1/hg38.tupChi1.synNet.maf.gz # 710111073 Apr 9 2015 mafLinks/mm10/hg38.mm10.synNet.maf.gz # 939260340 Apr 28 2015 mafLinks/cavPor3/hg38.cavPor3.synNet.maf.gz # 1144172783 Apr 26 2016 mafLinks/galVar1/hg38.galVar1.synNet.maf.gz # split the maf files into a set of hashed named files # this hash named split keeps the same chr/contig names in the same # named hash file. mkdir /hive/data/genomes/hg38/bed/multiz5way/mafSplit cd /hive/data/genomes/hg38/bed/multiz5way/mafSplit time for D in `sed -e "s/hg38 //" ../species.list` do echo "${D}" mkdir $D cd $D echo "mafSplit -byTarget -useHashedName=8 /dev/null . ../../mafLinks/${D}/*.maf.gz" mafSplit -byTarget -useHashedName=8 /dev/null . \ ../../mafLinks/${D}/*.maf.gz cd .. done # real 2m52.152s # construct a list of all possible maf file names. # they do not all exist in each of the species directories find . -type f | wc -l # 409 find . -type f | grep ".maf$" | xargs -L 1 basename | sort -u > maf.list wc -l maf.list # 108 maf.list mkdir /hive/data/genomes/hg38/bed/multiz5way/splitRun cd /hive/data/genomes/hg38/bed/multiz5way/splitRun mkdir maf run cd run mkdir penn cp -p /cluster/bin/penn/multiz.2009-01-21_patched/multiz penn cp -p /cluster/bin/penn/multiz.2009-01-21_patched/maf_project penn cp -p /cluster/bin/penn/multiz.2009-01-21_patched/autoMZ penn # verify the db and pairs settings are correct printf '#!/bin/csh -ef set db = hg38 set c = $1 set result = $2 set run = `/bin/pwd` set tmp = /dev/shm/$db/multiz.$c set pairs = /hive/data/genomes/hg38/bed/multiz5way/mafSplit /bin/rm -fr $tmp /bin/mkdir -p $tmp /bin/cp -p ../../tree.nh ../../species.list $tmp pushd $tmp > /dev/null foreach s (`/bin/sed -e "s/$db //" species.list`) set in = $pairs/$s/$c set out = $db.$s.sing.maf if (-e $in.gz) then /bin/zcat $in.gz > $out if (! -s $out) then echo "##maf version=1 scoring=autoMZ" > $out endif else if (-e $in) then /bin/ln -s $in $out else echo "##maf version=1 scoring=autoMZ" > $out endif end set path = ($run/penn $path); rehash $run/penn/autoMZ + T=$tmp E=$db "`cat tree.nh`" $db.*.sing.maf $c \ > /dev/null popd > /dev/null /bin/rm -f $result /bin/cp -p $tmp/$c $result /bin/rm -fr $tmp ' > autoMultiz.csh chmod +x autoMultiz.csh printf '#LOOP ./autoMultiz.csh $(file1) {check out line+ /hive/data/genomes/hg38/bed/multiz5way/splitRun/maf/$(root1).maf} #ENDLOOP ' > template # << happy emacs ln -s ../../mafSplit/maf.list maf.list ssh ku cd /hive/data/genomes/hg38/bed/multiz5way/splitRun/run gensub2 maf.list single template jobList para create jobList para try ... check ... push ... etc... # Completed: 108 of 108 jobs # CPU time in finished jobs: 69577s 1159.61m 19.33h 0.81d 0.002 y # IO & Wait Time: 409s 6.82m 0.11h 0.00d 0.000 y # Average job time: 648s 10.80m 0.18h 0.01d # Longest finished job: 6028s 100.47m 1.67h 0.07d # Submission to last job: 6042s 100.70m 1.68h 0.07d # combine into one file (the 1>&2 redirect sends the echo to stderr) cd /hive/data/genomes/hg38/bed/multiz5way head -1 splitRun/maf/006.maf > multiz5way.maf time for F in splitRun/maf/*.maf do echo "${F}" 1>&2 egrep -v "^#" ${F} done >> multiz5way.maf # real 0m31.596s tail -1 splitRun/maf/006.maf >> multiz5way.maf # -rw-rw-r-- 1 10395239286 Dec 17 21:03 multiz5way.maf # Load into database ssh hgwdev cd /hive/data/genomes/hg38/bed/multiz5way mkdir /gbdb/hg38/multiz5way ln -s `pwd`/multiz5way.maf /gbdb/hg38/multiz5way cd /dev/shm time hgLoadMaf hg38 multiz5way # Loaded 9874234 mafs in 1 files from /gbdb/hg38/multiz5way # real 2m55.355s time hgLoadMafSummary -verbose=2 -minSize=30000 \ -mergeGap=1500 -maxSize=200000 hg38 multiz5waySummary \ /gbdb/hg38/multiz5way/multiz5way.maf # Created 878615 summary blocks from 23435349 components and 9874234 mafs from /gbdb/hg38/multiz5way/multiz5way.maf # real 3m36.817s # -rw-rw-r-- 1 530340768 Dec 17 21:05 multiz5way.tab # -rw-rw-r-- 1 40949243 Dec 17 21:30 multiz5waySummary.tab wc -l multiz5way*.tab # 9874234 multiz5way.tab # 878615 multiz5waySummary.tab rm multiz5way*.tab ############################################################################## # GAP ANNOTATE MULTIZ7WAY MAF AND LOAD TABLES (DONE - 2017-12-17 - Hiram) # mafAddIRows has to be run on single chromosome maf files, it does not # function correctly when more than one reference sequence # are in a single file. Need to split of the maf file into individual # maf files mkdir -p /hive/data/genomes/hg38/bed/multiz5way/anno/mafSplit cd /hive/data/genomes/hg38/bed/multiz5way/anno/mafSplit time mafSplit -outDirDepth=2 -byTarget -useFullSequenceName \ /dev/null . ../../multiz5way.maf # real 2m57.457s find . -type f | wc -l # 332 # check for N.bed files everywhere: cd /hive/data/genomes/hg38/bed/multiz5way/anno for DB in `cat ../species.list` do if [ ! -s /hive/data/genomes/${DB}/${DB}.N.bed ]; then echo "MISS: ${DB}" # cd /hive/data/genomes/${DB} # twoBitInfo -nBed ${DB}.2bit ${DB}.N.bed else echo " OK: ${DB}" fi done cd /hive/data/genomes/hg38/bed/multiz5way/anno for DB in `cat ../species.list` do echo "${DB} " ln -s /hive/data/genomes/${DB}/${DB}.N.bed ${DB}.bed echo ${DB}.bed >> nBeds ln -s /hive/data/genomes/${DB}/chrom.sizes ${DB}.len echo ${DB}.len >> sizes done # make sure they all are successful symLinks: ls -ogrtL screen -S gapAnno # use a screen to control this longish job ssh ku cd /hive/data/genomes/hg38/bed/multiz5way/anno mkdir result find ./mafSplit -type d | sed -e 's#./mafSplit/##; s#./mafSplit##;' \ | while read D do echo mkdir -p result/${D} mkdir -p result/${D} done printf '#LOOP mafAddIRows -nBeds=nBeds mafSplit/$(path1) /hive/data/genomes/hg38/hg38.2bit {check out exists+ result/$(path1)} #ENDLOOP ' > template find ./mafSplit -type f | sed -e 's#^./mafSplit/##' > maf.list gensub2 maf.list single template jobList # limit jobs on a node with the ram=32g requirement because they go fast para -ram=32g create jobList para try ... check ... push ... # Completed: 332 of 332 jobs # CPU time in finished jobs: 717s 11.95m 0.20h 0.01d 0.000 y # IO & Wait Time: 906s 15.10m 0.25h 0.01d 0.000 y # Average job time: 5s 0.08m 0.00h 0.00d # Longest finished job: 62s 1.03m 0.02h 0.00d # Submission to last job: 96s 1.60m 0.03h 0.00d # verify all result files have some content, look for 0 size files: find ./result -type f -size 0 # should see none # or in this manner: find ./result -type f | xargs ls -og | sort -k3nr | tail # combine into one file (the 1>&2 redirect sends the echo to stderr) head -q -n 1 result/0/0/chr14_KI270845v1_alt.maf > hg38.5way.maf time find ./result -type f | while read F do echo "${F}" 1>&2 grep -h -v "^#" ${F} done >> hg38.5way.maf # real 0m43.949s # these maf files do not have the end marker, this does nothing: # tail -q -n 1 result/0/0/chr14_KI270845v1_alt.maf >> hg38.5way.maf # How about an official end marker: echo "##eof maf" >> hg38.5way.maf ls -og # -rw-rw-r-- 1 13290077466 Dec 17 22:37 hg38.5way.maf du -hsc hg38.5way.maf ../*.maf # 13G hg38.5way.maf # 9.7G ../multiz5way.maf # construct symlinks to get the individual maf files into gbdb: rm /gbdb/hg38/multiz5way/multiz5way.maf # remove previous results ln -s `pwd`/hg38.5way.maf /gbdb/hg38/multiz5way/multiz5way.maf # Load into database cd /dev/shm time hgLoadMaf -pathPrefix=/gbdb/hg38/multiz5way hg38 multiz5way # Loaded 11023447 mafs in 1 files from /gbdb/hg38/multiz5way # real 3m37.392s time hgLoadMafSummary -verbose=2 -minSize=30000 \ -mergeGap=1500 -maxSize=200000 hg38 multiz5waySummary \ /gbdb/hg38/multiz5way/multiz5way.maf # Created 878615 summary blocks from 23435349 components and 11023447 mafs from /gbdb/hg38/multiz5way/multiz5way.maf # real 4m10.706s # -rw-rw-r-- 1 594646492 Dec 17 22:41 multiz5way.tab # -rw-rw-r-- 1 42706473 Dec 17 22:48 multiz5waySummary.tab rm multiz5way*.tab ###################################################################### # MULTIZ7WAY MAF FRAMES (TBD - 2016-05-05 - Hiram) ssh hgwdev mkdir /hive/data/genomes/hg38/bed/multiz5way/frames cd /hive/data/genomes/hg38/bed/multiz5way/frames # survey all the genomes to find out what kinds of gene tracks they have printf '#!/bin/csh -fe foreach db (`cat ../species.list`) printf "# ${db}: " set tables = `hgsql $db -N -e "show tables" | egrep "Gene|ncbiRefSeq"` foreach table ($tables) if ($table == "ensGene" || $table == "refGene" || \ $table == "ncbiRefSeq" || $table == "mgcGenes" || \ $table == "knownGene" || $table == "xenoRefGene" ) then set count = `hgsql $db -N -e "select count(*) from $table"` echo -n "${table}: ${count}, " endif end set orgName = `hgsql hgcentraltest -N -e \ "select scientificName from dbDb where name='"'"'$db'"'"'"` set orgId = `hgsql hgFixed -N -e \ "select id from organism where name='"'"'$orgName'"'"'"` if ($orgId == "") then echo "Mrnas: 0" else set count = `hgsql hgFixed -N -e "select count(*) from gbCdnaInfo where organism=$orgId"` echo "Mrnas: ${count}" endif end ' > showGenes.csh # << happy emacs chmod +x ./showGenes.csh time ./showGenes.csh # hg38: ensGene: 208239, knownGene: 196838, mgcGenes: 35312, ncbiRefSeq: 159322, refGene: 74453, xenoRefGene: 187376, Mrnas: 11508577 # tupChi1: refGene: 206, xenoRefGene: 353563, Mrnas: 50709 # galVar1: ncbiRefSeq: 41547, xenoRefGene: 516902, Mrnas: 0 # mm10: ensGene: 103734, knownGene: 63759, mgcGenes: 27612, ncbiRefSeq: 106520, refGene: 39240, xenoRefGene: 183459, Mrnas: 5371140 # cavPor3: ensGene: 34846, refGene: 488, xenoRefGene: 316945, Mrnas: 21241 # real 1m5.600s # from that summary, use these gene sets: # knownGene - hg38 mm10 # ensGene - cavPor3 # none - galVar1 tupChi1 mkdir genes # 1. knownGene: hg38 mm10 for DB in hg38 mm10 do hgsql -N -e "select name,chrom,strand,txStart,txEnd,cdsStart,cdsEnd,exonCount,exonStarts,exonEnds from knownGene" ${DB} \ | genePredSingleCover stdin stdout | gzip -2c \ > genes/${DB}.gp.gz printf "# ${DB}: " genePredCheck -db=${DB} genes/${DB}.gp.gz done # hg38: checked: 21554 failed: 0 # mm10: checked: 21100 failed: 0 # 2. ensGene: cavPor3 for DB in cavPor3 do hgsql -N -e "select name,chrom,strand,txStart,txEnd,cdsStart,cdsEnd,exonCount,exonStarts,exonEnds from ensGene" ${DB} \ | genePredSingleCover stdin stdout | gzip -2c \ > /dev/shm/${DB}.tmp.gz mv /dev/shm/${DB}.tmp.gz genes/$DB.gp.gz printf "# ${DB}: " genePredCheck -db=${DB} genes/${DB}.gp.gz done # cavPor3: checked: 18034 failed: 0 # verify counts for genes are reasonable: for T in genes/*.gz do echo -n "# $T: " zcat $T | cut -f1 | sort | uniq -c | wc -l done # genes/cavPor3.gp.gz: 18034 # genes/hg38.gp.gz: 21554 # genes/mm10.gp.gz: 21100 time (cat ../anno/hg38.5way.maf \ | genePredToMafFrames hg38 stdin stdout \ `sed -e 's/tupChi1//; s/galVar1//;' ../species.list.txt | xargs echo \ | sed -e "s#\([a-zA-Z0-9]*\)#\1 genes/\1.gp.gz#g;"` \ | gzip > multiz5wayFrames.bed.gz) # real 2m39.249s # verify there are frames on everything, should be 3 species: zcat multiz5wayFrames.bed.gz | awk '{print $4}' | sort | uniq -c \ | sed -e 's/^/# /;' # 241239 cavPor3 # 206671 hg38 # 261120 mm10 # load the resulting file ssh hgwdev cd /hive/data/genomes/hg38/bed/multiz5way/frames time hgLoadMafFrames hg38 multiz5wayFrames multiz5wayFrames.bed.gz # real 0m7.413s time featureBits -countGaps hg38 multiz5wayFrames # 39254551 bases of 3209286105 (1.223%) in intersection # real 0m5.378s # enable the trackDb entries: # frames multiz5wayFrames # irows on # appears to work OK ######################################################################### # Phylogenetic tree from 5-way (TBD - 2016-05-06 - Hiram) mkdir /hive/data/genomes/hg38/bed/multiz5way/4d cd /hive/data/genomes/hg38/bed/multiz5way/4d # using knownGene for hg38, only transcribed genes and nothing # from the randoms and other misc. hgsql -Ne "select name,chrom,strand,txStart,txEnd,cdsStart,cdsEnd,exonCount,exonStarts,exonEnds from knownGene where cdsEnd > cdsStart;" hg38 \ | egrep -E -v "chrM|chrUn|random|_alt" \ | genePredSingleCover stdin stdout > hg38.knownGeneNR.gp genePredCheck -db=hg38 hg38.knownGeneNR.gp # checked: 19306 failed: 0 # the annotated maf is: og ../anno/hg38.5way.maf # -rw-rw-r-- 1 13290077466 Dec 17 22:37 ../anno/hg38.5way.maf mkdir annoSplit cd annoSplit time mafSplit -verbose=2 -outDirDepth=2 -byTarget -useFullSequenceName \ /dev/null . ../../anno/hg38.5way.maf # real 4m9.934s find . -type f | wc -l # 332 ssh ku mkdir /hive/data/genomes/hg38/bed/multiz5way/4d/run cd /hive/data/genomes/hg38/bed/multiz5way/4d/run mkdir ../mfa cat << '_EOF_' > 4d.csh #!/bin/csh -fex set PHASTBIN = /cluster/bin/phast.build/cornellCVS/phast.2010-12-30/bin set GP = hg38.knownGeneNR.gp set r = "/hive/data/genomes/hg38/bed/multiz5way" set c = $1 set infile = $r/4d/$2 set outDir = $r/4d/$3 set outfile = $r/4d/run/$4 /bin/mkdir -p $outDir cd /dev/shm /bin/awk -v C=$c '$2 == C {print}' $r/4d/$GP | sed -e "s/\t$c\t/\ttupChi1.$c\t/" > $c.gp set NL=`wc -l $c.gp| gawk '{print $1}'` echo $NL if ("$NL" != "0") then $PHASTBIN/msa_view --4d --features $c.gp -i MAF $infile -o SS > $c.ss $PHASTBIN/msa_view -i SS --tuple-size 1 $c.ss > $outfile else echo "" > $outfile endif /bin/rm -f /dev/shm/$c.gp /dev/shm/$c.ss _EOF_ # << happy emacs chmod +x 4d.csh find ../annoSplit -type f | sed -e "s#../annoSplit/##" > maf.list wc -l maf.list # 332 maf.list printf '#LOOP 4d.csh $(root1) annoSplit/$(dir1)/$(file1) mfa/$(dir1) {check out line+ ../mfa/$(dir1)/$(root1).mfa} #ENDLOOP ' > template gensub2 maf.list single template jobList para create jobList para try ... check para time # Completed: 330 of 332 jobs # Crashed: 2 jobs # CPU time in finished jobs: 585s 9.76m 0.16h 0.01d 0.000 y # IO & Wait Time: 773s 12.88m 0.21h 0.01d 0.000 y # Average job time: 4s 0.07m 0.00h 0.00d # Longest finished job: 54s 0.90m 0.01h 0.00d # Submission to last job: 91s 1.52m 0.03h 0.00d # Not all results have contents, or finish successfully, that is OK # it is because not all contigs have genes, only gene sequences are measured # combine mfa files ssh hgwdev cd /hive/data/genomes/hg38/bed/multiz5way/4d # remove the broken empty files, size 0 and size 1: find ./mfa -type f -size 0 | xargs rm -f # sometimes this doesn't work, don't know why ### XXX does not work find ./mfa -type f -size 1 | xargs rm -f # when it doesn't, use this empty list procedure find ./mfa -type f | xargs ls -og | awk '$3 < 2' | awk '{print $NF}' \ > empty.list cat empty.list | xargs rm -f # see what is left: ls -ogrt mfa/*/*/*.mfa | sort -k3nr | wc # 22 154 1091 # want comma-less species.list time /cluster/bin/phast.build/cornellCVS/phast.2010-12-30/bin/msa_view \ --aggregate "`cat ../species.list`" mfa/*/*/*.mfa | sed s/"> "/">"/ \ > 4d.all.mfa # real 0m0.426s # check they are all in there: grep "^>" 4d.all.mfa | wc -l # 5 grep "^>" 4d.all.mfa | sed -e 's/^/# /;' # >hg38 # >tupChi1 # >galVar1 # >mm10 # >cavPor3 sed 's/[a-z][a-z]*_//g; s/:[0-9\.][0-9\.]*//g; s/;//; /^ *$/d' \ ../hg38.5way.nh | xargs echo | sed -e 's/ //g' > tree_commas.nh # tree_commas.nh looks like: # ((hg38,(tupChi1,galVar1)),(mm10,cavPor3)) # use phyloFit to create tree model (output is phyloFit.mod) time /cluster/bin/phast.build/cornellCVS/phast.2010-12-30/bin/phyloFit \ --EM --precision MED --msa-format FASTA --subst-mod REV \ --tree tree_commas.nh 4d.all.mfa # real 0m0.256s mv phyloFit.mod all.mod grep TREE all.mod # TREE: # ((hg38:0.0700315,(tupChi1:0.0981977,galVar1:0.0629139):0.00881734):0.0161669, # (mm10:0.142138,cavPor3:0.112136):0.0161669); # compare these calculated lengths to the tree extracted from 218way: grep TREE all.mod | sed -e 's/TREE: //' \ | /cluster/bin/phast/all_dists /dev/stdin | grep hg38 \ | sed -e "s/hg38.//;" | sort > new.dists /cluster/bin/phast/all_dists ../hg38.5way.nh | grep hg38 \ | sed -e "s/hg38.//;" | sort > old.dists # printing out the 'new', the 'old' the 'difference' and percent difference join new.dists old.dists | awk '{ printf "#\t%s\t%8.6f\t%8.6f\t%8.6f\t%8.6f\n", $1, $2, $3, $2-$3, 100*($2-$3)/$3 }' \ | sort -k3n # galVar1 0.141763 0.278845 -0.137082 -49.160645 # tupChi1 0.177047 0.318845 -0.141798 -44.472393 # cavPor3 0.214501 0.362746 -0.148245 -40.867439 # mm10 0.244503 0.502391 -0.257888 -51.332130 XXX - ready to continue - Mon Dec 18 11:34:03 PST 2017 ######################################################################### # phastCons 5-way (TBD - 2016-05-06 - Hiram) # split 5way mafs into 10M chunks and generate sufficient statistics # files for # phastCons ssh ku mkdir -p /hive/data/genomes/hg38/bed/multiz5way/cons/SS cd /hive/data/genomes/hg38/bed/multiz5way/cons/SS mkdir result done printf '#!/bin/csh -ef set d = $1 set c = $2 set doneDir = done/$d set MAF = /hive/data/genomes/hg38/bed/multiz5way/anno/result/$d/$c.maf set WINDOWS = /hive/data/genomes/hg38/bed/multiz5way/cons/SS/result/$d/$c set WC = `cat $MAF | wc -l` set NL = `grep "^#" $MAF | wc -l` if ( -s $3 ) then exit 0 endif if ( -s $3.running ) then exit 0 endif /bin/mkdir -p $doneDir /bin/date >> $3.running /bin/rm -fr $WINDOWS /bin/mkdir -p $WINDOWS pushd $WINDOWS > /dev/null if ( $WC != $NL ) then /cluster/bin/phast.build/cornellCVS/phast.2010-12-30/bin/msa_split \\ $MAF -i MAF -o SS -r $WINDOWS/$c -w 10000000,0 -I 1000 -B 5000 endif popd > /dev/null /bin/date >> $3 /bin/rm -f $3.running ' > mkSS.csh chmod +x mkSS.csh printf '#LOOP mkSS.csh $(dir1) $(root1) {check out line+ done/$(dir1)/$(root1)} #ENDLOOP ' > template find ../../anno/result -type f | sed -e "s#../../anno/result/##" > maf.list wc -l maf.list # 76237 maf.list ssh ku cd /hive/data/genomes/hg38/bed/multiz5way/cons/SS gensub2 maf.list single template jobList # beware overwhelming the cluster with these quick high I/O jobs para create jobList para try ... check ... etc para -maxJob=64 push # Completed: 76237 of 76237 jobs # CPU time in finished jobs: 3491s 58.19m 0.97h 0.04d 0.000 y # IO & Wait Time: 321255s 5354.25m 89.24h 3.72d 0.010 y # Average job time: 4s 0.07m 0.00h 0.00d # Longest finished job: 10s 0.17m 0.00h 0.00d # Submission to last job: 1583s 26.38m 0.44h 0.02d find ./result -type f | wc -l # 24853 # Run phastCons # This job is I/O intensive in its output files, beware where this # takes place or do not run too many at once. ssh ku mkdir -p /hive/data/genomes/hg38/bed/multiz5way/cons/run.cons cd /hive/data/genomes/hg38/bed/multiz5way/cons/run.cons # This is setup for multiple runs based on subsets, but only running # the 'all' subset here. # It triggers off of the current working directory # $cwd:t which is the "grp" in this script. Running: # all and vertebrates printf '#!/bin/csh -fe set PHASTBIN = /cluster/bin/phast.build/cornellCVS/phast.2010-12-30/bin set c = $1 set d = $2 set f = $3 set len = $4 set cov = $5 set rho = $6 set grp = $cwd:t set cons = /hive/data/genomes/hg38/bed/multiz5way/cons set tmp = $cons/tmp/${d}_${c} mkdir -p $tmp set ssSrc = $cons/SS/result set useGrp = "$grp.mod" if (-s $cons/$grp/$grp.non-inf) then ln -s $cons/$grp/$grp.mod $tmp ln -s $cons/$grp/$grp.non-inf $tmp ln -s $ssSrc/$d/$f $tmp else ln -s $ssSrc/$d/$f $tmp ln -s $cons/$grp/$grp.mod $tmp endif pushd $tmp > /dev/null if (-s $grp.non-inf) then $PHASTBIN/phastCons $f $useGrp \ --rho $rho --expected-length $len --target-coverage $cov --quiet \\ --not-informative `cat $grp.non-inf` \\ --seqname $c --idpref $c --most-conserved $c.bed --score > $c.pp else $PHASTBIN/phastCons $f $useGrp \\ --rho $rho --expected-length $len --target-coverage $cov --quiet \\ --seqname $c --idpref $c --most-conserved $c.bed --score > $c.pp endif popd > /dev/null mkdir -p pp/$d bed/$d sleep 4 touch pp/$d bed/$d rm -f pp/$d/$c.pp rm -f bed/$d/$c.bed mv $tmp/$c.pp pp/$d mv $tmp/$c.bed bed/$d rm -fr $tmp rmdir --ignore-fail-on-non-empty $cons/tmp/$d:h ' > doPhast.csh chmod +x doPhast.csh # this template will serve for all runs # root1 == chrom name, file1 == ss file name without .ss suffix printf '#LOOP ../run.cons/doPhast.csh $(root1) $(dir1) $(file1) 45 0.3 0.3 {check out line+ pp/$(dir1)/$(root1).pp} #ENDLOOP ' > template find ../SS/result -type f | sed -e "s#../SS/result/##" > ss.list wc -l ss.list # 24853 ss.list # Create parasol batch and run it # run for all species cd /hive/data/genomes/hg38/bed/multiz5way/cons mkdir -p all cd all # Using the .mod tree cp -p ../../4d/all.mod ./all.mod gensub2 ../run.cons/ss.list single ../run.cons/template jobList para -ram=32g create jobList para try ... check ... para push # Completed: 24815 of 24853 jobs # Crashed: 38 jobs # CPU time in finished jobs: 6246s 104.09m 1.73h 0.07d 0.000 y # IO & Wait Time: 222621s 3710.36m 61.84h 2.58d 0.007 y # Average job time: 9s 0.15m 0.00h 0.00d # Longest finished job: 18s 0.30m 0.01h 0.00d # Submission to last job: 935s 15.58m 0.26h 0.01d # the 38 crash jobs were actually finished, they failed the last rmdir: # rmdir: failed to remove `/hive/data/genomes/hg38/bed/multiz5way/cons/tmp/7/6': No such file or directory # create Most Conserved track cd /hive/data/genomes/hg38/bed/multiz5way/cons/all time cut -f1 ../../../../chrom.sizes | while read C do ls -d bed/?/?/${C} 2> /dev/null | while read D do echo ${D}/${C}*.bed 1>&2 cat ${D}/${C}*.bed done | sort -k1,1 -k2,2n \ | awk '{printf "%s\t%d\t%d\tlod=%d\t%s\n", "'${C}'", $2, $3, $5, $5;}' done > tmpMostConserved.bed # real 19m26.845s time /cluster/bin/scripts/lodToBedScore tmpMostConserved.bed \ > mostConserved.bed # real 0m5.667s # -rw-rw-r-- 1 36626033 May 6 14:55 mostConserved.bed # load into database ssh hgwdev cd /hive/data/genomes/hg38/bed/multiz5way/cons/all time hgLoadBed hg38 phastConsElements5way mostConserved.bed # Read 932856 elements of size 5 from mostConserved.bed # real 0m9.898s # on human we often try for 5% overall cov, and 70% CDS cov # most bets are off here for that goal, these alignments are too few # and too far between # --rho 0.3 --expected-length 45 --target-coverage 0.3 time featureBits hg38 -enrichment ncbiRefSeq:cds phastConsElements5way # ncbiRefSeq:cds 1.217%, phastConsElements5way 3.976%, both 0.819%, # cover 67.27%, enrich 16.92x # real 2m33.330s # Create merged posterier probability file and wiggle track data files cd /hive/data/genomes/hg38/bed/multiz5way/cons/all mkdir downloads # the third sed fixes the chrom names, removing the partition extensions time (find ./pp -type f | sed -e "s#^./##; s#\.# d #g; s#-# m #;" \ | sort -k1,1 -k3,3n | sed -e "s# d #.#g; s# m #-#g;" | xargs cat \ | sed -e 's/\.[0-9][0-9]*-[0-9][0-9]* start/ start/' \ | gzip -c > downloads/phastCons5way.wigFix.gz) # real 27m47.145s # -rw-rw-r-- 1 2207346680 May 6 15:33 phastCons5way.wigFix.gz # check integrity of data with wigToBigWig time (zcat downloads/phastCons5way.wigFix.gz \ | wigToBigWig -verbose=2 stdin /hive/data/genomes/hg38/chrom.sizes \ phastCons5way.bw) > bigWig.log 2>&1 egrep "real|VmPeak" bigWig.log # pid=37326: VmPeak: 20943944 kB # real 30m30.283s bigWigInfo phastCons5way.bw | sed -e 's/^/# /;' # version: 4 # isCompressed: yes # isSwapped: 0 # primaryDataSize: 3,364,371,979 # primaryIndexSize: 81,959,008 # zoomLevels: 10 # chromCount: 24853 # basesCovered: 1,910,088,593 # mean: 0.117235 # min: 0.000000 # max: 1.000000 # std: 0.237776 # encode those files into wiggle data time (zcat downloads/phastCons5way.wigFix.gz \ | wigEncode stdin phastCons5way.wig phastCons5way.wib) # Converted stdin, upper limit 1.00, lower limit 0.00 # real 10m31.797s du -hsc *.wi? # 1.8G phastCons5way.wib # 275M phastCons5way.wig # Load gbdb and database with wiggle. ln -s `pwd`/phastCons5way.wib /gbdb/hg38/multiz5way/phastCons5way.wib time hgLoadWiggle -pathPrefix=/gbdb/hg38/multiz5way \ hg38 phastCons5way phastCons5way.wig # real 0m30.803s # use to set trackDb.ra entries for wiggle min and max # and verify table is loaded correctly wigTableStats.sh hg38 phastCons5way # db.table min max mean count sumData # hg38.phastCons5way 0 1 0.117235 1910088593 2.23929e+08 # stdDev viewLimits # 0.237776 viewLimits=0:1 # Create histogram to get an overview of all the data time hgWiggle -doHistogram -db=hg38 \ -hBinSize=0.001 -hBinCount=1000 -hMinVal=0.0 -verbose=2 \ phastCons5way > histogram.data 2>&1 # real 4m6.489s # create plot of histogram: printf 'set terminal png small x000000 xffffff xc000ff x66ff66 xffff00 x00ffff font \ "/usr/share/fonts/default/Type1/n022004l.pfb" set size 1.4, 0.8 set key left box set grid noxtics set grid ytics set title " Malayan flying lemur hg38 Histogram phastCons5way track" set xlabel " phastCons5way score" set ylabel " Relative Frequency" set y2label " Cumulative Relative Frequency (CRF)" set y2range [0:1] set y2tics set yrange [0:0.02] plot "histogram.data" using 2:5 title " RelFreq" with impulses, \\ "histogram.data" using 2:7 axes x1y2 title " CRF" with lines ' | gnuplot > histo.png display histo.png & ######################################################################### # phyloP for 5-way (TBD - 2016-05-09,11 - Hiram) # run phyloP with score=LRT ssh ku mkdir /cluster/data/hg38/bed/multiz5way/consPhyloP cd /cluster/data/hg38/bed/multiz5way/consPhyloP mkdir run.phyloP cd run.phyloP # Adjust model file base composition background and rate matrix to be # representative of the chromosomes in play grep BACKGROUND ../../4d/all.mod | awk '{printf "%0.3f\n", $3 + $4}' # 0.562 /cluster/bin/phast.build/cornellCVS/phast.2010-12-30/bin/modFreqs \ ../../4d/all.mod 0.562 > all.mod # verify, the BACKGROUND should now be paired up: grep BACK all.mod # BACKGROUND: 0.219000 0.281000 0.281000 0.219000 printf '#!/bin/csh -fe set PHASTBIN = /cluster/bin/phast.build/cornellCVS/phast.2010-12-30/bin set f = $1 set d = $f:h set file1 = $f:t set out = $2 set cName = $f:t:r set grp = $cwd:t set cons = /hive/data/genomes/hg38/bed/multiz5way/consPhyloP set tmp = $cons/tmp/$grp/$f /bin/rm -fr $tmp /bin/mkdir -p $tmp set ssSrc = "/hive/data/genomes/hg38/bed/multiz5way/cons/SS/result/$f" set useGrp = "$grp.mod" /bin/ln -s $cons/run.phyloP/$grp.mod $tmp pushd $tmp > /dev/null $PHASTBIN/phyloP --method LRT --mode CONACC --wig-scores --chrom $cName \\ -i SS $useGrp $ssSrc.ss > $file1.wigFix popd > /dev/null /bin/mkdir -p $out:h sleep 4 /bin/touch $out:h /bin/mv $tmp/$file1.wigFix $out /bin/rm -fr $tmp /bin/rmdir --ignore-fail-on-non-empty $cons/tmp/$grp/$d /bin/rmdir --ignore-fail-on-non-empty $cons/tmp/$grp/$d:h /bin/rmdir --ignore-fail-on-non-empty $cons/tmp/$grp /bin/rmdir --ignore-fail-on-non-empty $cons/tmp ' > doPhyloP.csh chmod +x doPhyloP.csh # Create list of chunks find ../../cons/SS/result -type f | grep ".ss$" \ | sed -e "s/.ss$//; s#^../../cons/SS/result/##" > ss.list # make sure the list looks good wc -l ss.list # 24853 ss.list # Create template file # file1 == $chr/$chunk/file name without .ss suffix printf '#LOOP ../run.phyloP/doPhyloP.csh $(path1) {check out line+ wigFix/$(dir1)/$(file1).wigFix} #ENDLOOP ' > template ###################### Running all species ####################### # setup run for all species mkdir /hive/data/genomes/hg38/bed/multiz5way/consPhyloP/all cd /hive/data/genomes/hg38/bed/multiz5way/consPhyloP/all rm -fr wigFix mkdir wigFix gensub2 ../run.phyloP/ss.list single ../run.phyloP/template jobList # beware overwhelming the cluster with these fast running high I/O jobs para create jobList para try ... check ... push ... etc ... para -maxJob=53 push para time > run.time # Completed: 24852 of 24853 jobs # Crashed: 1 jobs # CPU time in finished jobs: 7517s 125.29m 2.09h 0.09d 0.000 y # IO & Wait Time: 166287s 2771.45m 46.19h 1.92d 0.005 y # Average job time: 7s 0.12m 0.00h 0.00d # Longest finished job: 11s 0.18m 0.00h 0.00d # Submission to last job: 1799s 29.98m 0.50h 0.02d # the one failed job was just the last rmdir command: # /bin/rmdir: failed to remove `/hive/data/genomes/hg38/bed/multiz5way/consPhyloP/tmp/all/5/2 mkdir downloads time (find ./wigFix -type f | sed -e "s#^./##; s#\.# d #g; s#-# m #;" \ | sort -k1,1 -k3,3n | sed -e "s# d #.#g; s# m #-#g;" | xargs cat \ | gzip -c > downloads/phyloP5way.wigFix.gz) # real 30m20.572s # check integrity of data with wigToBigWig time (zcat downloads/phyloP5way.wigFix.gz \ | wigToBigWig -verbose=2 stdin /hive/data/genomes/hg38/chrom.sizes \ phyloP5way.bw) > bigWig.log 2>&1 egrep "real|VmPeak" bigWig.log # pid=19896: VmPeak: 20943916 kB # real 215m36.588s bigWigInfo phyloP5way.bw | sed -e 's/^/# /;' # version: 4 # isCompressed: yes # isSwapped: 0 # primaryDataSize: 3,067,676,886 # primaryIndexSize: 81,959,008 # zoomLevels: 10 # chromCount: 24853 # basesCovered: 1,910,088,593 # mean: 0.061208 # min: -2.599000 # max: 0.833000 # std: 0.611659 # encode those files into wiggle data time (zcat downloads/phyloP5way.wigFix.gz \ | wigEncode stdin phyloP5way.wig phyloP5way.wib) # Converted stdin, upper limit 0.83, lower limit -2.60 # real 9m4.543s du -hsc *.wi? # 1.8G phyloP5way.wib # 279M phyloP5way.wig # Load gbdb and database with wiggle. ln -s `pwd`/phyloP5way.wib /gbdb/hg38/multiz5way/phyloP5way.wib time hgLoadWiggle -pathPrefix=/gbdb/hg38/multiz5way hg38 \ phyloP5way phyloP5way.wig # real 0m30.869s # use to set trackDb.ra entries for wiggle min and max # and verify table is loaded correctly wigTableStats.sh hg38 phyloP5way # db.table min max mean count sumData # hg38.phyloP5way -2.599 0.833 0.0612084 1910088593 1.16914e+08 # stdDev viewLimits # 0.611659 viewLimits=-2.599:0.833 # that range is: 0.833+2.599 = 3.432 for hBinSize=0.003432 # Create histogram to get an overview of all the data time hgWiggle -doHistogram \ -hBinSize=0.003432 -hBinCount=1000 -hMinVal=-2.559 -verbose=2 \ -db=hg38 phyloP5way > histogram.data 2>&1 # real 4m20.444s # find the Y range for the 2:5 graph grep -v chrom histogram.data | grep "^[0-9]" | ave -col=5 stdin \ | sed -e 's/^/# /;' # Q1 0.000087 # median 0.000381 # Q3 0.001361 # average 0.001112 # min 0.000000 # max 0.032088 # count 899 # total 0.999990 # standard deviation 0.002165 # find the X range for the 2:5 graph grep "^[0-9]" histogram.data | ave -col=2 stdin \ | sed -e 's/^/# /;' # Q1 -1.773070 # median -1.004300 # Q3 -0.228672 # average -0.962349 # min -2.559000 # max 0.831816 # count 899 # total -865.151612 # standard deviation 0.954068 # create plot of histogram: printf 'set terminal png small x000000 xffffff xc000ff x66ff66 xffff00 x00ffff font \ "/usr/share/fonts/default/Type1/n022004l.pfb" set size 1.4, 0.8 set key left box set grid noxtics set grid ytics set title " Guinea pig hg38 Histogram phyloP5way track" set xlabel " phyloP5way score" set ylabel " Relative Frequency" set y2label " Cumulative Relative Frequency (CRF)" set y2range [0:1] set y2tics set xtics set xrange [-2.6:0.85] set yrange [0:0.033] plot "histogram.data" using 2:5 title " RelFreq" with impulses, \ "histogram.data" using 2:7 axes x1y2 title " CRF" with lines ' | gnuplot > histo.png display histo.png & # appears to have an odd hole in the data near X=0 ? ############################################################################# # hgPal downloads (TBD - 2016-05-09,11 - Hiram) # FASTA from 5-way for knownGene, refGene and knownCanonical ssh hgwdev screen -S hg38HgPal mkdir /hive/data/genomes/hg38/bed/multiz5way/pal cd /hive/data/genomes/hg38/bed/multiz5way/pal cat ../species.list | tr '[ ]' '[\n]' > order.list # this for loop takes about 2.5 hours on this large count contig assembly export mz=multiz5way export gp=ncbiRefSeq export db=hg38 export I=0 export D=0 mkdir exonAA exonNuc printf '#!/bin/sh\n' > $gp.jobs time for C in `sort -nk2 ../../../chrom.sizes | cut -f1` do I=`echo $I | awk '{print $1+1}'` D=`echo $D | awk '{print $1+1}'` dNum=`echo $D | awk '{printf "%03d", int($1/1000)}'` mkdir -p exonNuc/${dNum} > /dev/null mkdir -p exonAA/${dNum} > /dev/null echo "mafGene -chrom=$C -exons -noTrans $db $mz $gp order.list stdout | gzip -c > exonNuc/${dNum}/$C.exonNuc.fa.gz &" echo "mafGene -chrom=$C -exons $db $mz $gp order.list stdout | gzip -c > exonAA/${dNum}/$C.exonAA.fa.gz &" if [ $I -gt 16 ]; then echo "date" echo "wait" I=0 fi done >> $gp.jobs # real 115m16.333s echo "date" >> $gp.jobs echo "wait" >> $gp.jobs chmod +x ncbiRefSeq.jobs time (./$gp.jobs) > $gp.jobs.log 2>&1 & # real 14m50.750s export mz=multiz5way export gp=ncbiRefSeq time find ./exonAA -type f | grep exonAA.fa.gz | xargs zcat \ | gzip -c > $gp.$mz.exonAA.fa.gz # real 4m20.026s time find ./exonNuc -type f | grep exonNuc.fa.gz | xargs zcat \ | gzip -c > $gp.$mz.exonNuc.fa.gz # real 4m45.761s # -rw-rw-r-- 1 36201970 May 11 11:25 ncbiRefSeq.multiz5way.exonAA.fa.gz # -rw-rw-r-- 1 59404213 May 11 11:30 ncbiRefSeq.multiz5way.exonNuc.fa.gz export mz=multiz5way export gp=ncbiRefSeq export db=hg38 export pd=/usr/local/apache/htdocs-hgdownload/goldenPath/$db/$mz/alignments mkdir -p $pd md5sum *.fa.gz > md5sum.txt ln -s `pwd`/$gp.$mz.exonAA.fa.gz $pd/$gp.exonAA.fa.gz ln -s `pwd`/$gp.$mz.exonNuc.fa.gz $pd/$gp.exonNuc.fa.gz ln -s `pwd`/md5sum.txt $pd/ rm -rf exonAA exonNuc ############################################################################# # construct download files for 5-way (TBD - 2016-05-11 - Hiram) mkdir /usr/local/apache/htdocs-hgdownload/goldenPath/hg38/multiz5way mkdir /usr/local/apache/htdocs-hgdownload/goldenPath/hg38/phastCons5way mkdir /usr/local/apache/htdocs-hgdownload/goldenPath/hg38/phyloP5way mkdir /hive/data/genomes/hg38/bed/multiz5way/downloads cd /hive/data/genomes/hg38/bed/multiz5way/downloads mkdir multiz5way phastCons5way phyloP5way cd multiz5way time cp -p ../../anno/hg38.5way.maf . # real 0m20.564s # -rw-rw-r-- 1 13290077466 Dec 17 22:37 hg38.5way.maf du -hsc * # 13G hg38.5way.maf time gzip *.maf # real 43m14.605s # -rw-rw-r-- 1 3422259860 Dec 17 22:37 hg38.5way.maf.gz du -hsc *.maf.gz # 3.2G hg38.5way.maf.gz grep TREE ../../4d/all.mod | awk '{print $NF}' \ | ~/kent/src/hg/utils/phyloTrees/asciiTree.pl /dev/stdin \ > hg38.5way.nh ~/kent/src/hg/utils/phyloTrees/commonNames.sh hg38.5way.nh \ | ~/kent/src/hg/utils/phyloTrees/asciiTree.pl /dev/stdin \ > hg38.5way.commonNames.nh ~/kent/src/hg/utils/phyloTrees/scientificNames.sh hg38.5way.nh \ | $HOME/kent/src/hg/utils/phyloTrees/asciiTree.pl /dev/stdin \ > hg38.5way.scientificNames.nh time md5sum *.nh *.maf.gz > md5sum.txt # real 0m35.144s ln -s `pwd`/* \ /usr/local/apache/htdocs-hgdownload/goldenPath/hg38/multiz5way du -hsc *.maf.gz ../../anno/hg38.5way.maf # 3.2G hg38.5way.maf.gz # 13G ../../anno/hg38.5way.maf XXX - writing README - Mon Dec 18 11:41:15 PST 2017 # obtain the README.txt from cavPor3/multiz5way and update for this # situation ##################################################################### cd /hive/data/genomes/hg38/bed/multiz5way/downloads/phastCons5way ln -s ../../cons/all/downloads/phastCons5way.wigFix.gz \ ./hg38.phastCons5way.wigFix.gz ln -s ../../cons/all/phastCons5way.bw ./hg38.phastCons5way.bw ln -s ../../cons/all/all.mod ./hg38.phastCons5way.mod time md5sum *.gz *.mod *.bw > md5sum.txt # real 0m20.354s # obtain the README.txt from cavPor3/phastCons5way and update for this # situation ln -s `pwd`/* \ /usr/local/apache/htdocs-hgdownload/goldenPath/hg38/phastCons5way ##################################################################### cd /hive/data/genomes/hg38/bed/multiz5way/downloads/phyloP5way ln -s ../../consPhyloP/all/downloads/phyloP5way.wigFix.gz \ ./hg38.phyloP5way.wigFix.gz ln -s ../../consPhyloP/run.phyloP/all.mod hg38.phyloP5way.mod ln -s ../../consPhyloP/all/phyloP5way.bw hg38.phyloP5way.bw time md5sum *.mod *.bw *.gz > md5sum.txt # real 0m29.662s # obtain the README.txt from hg38/phyloP17way and update for this # situation ln -s `pwd`/* \ /usr/local/apache/htdocs-hgdownload/goldenPath/hg38/phyloP5way ########################################################################### ## create upstream refGene maf files cd /hive/data/genomes/hg38/bed/multiz5way/downloads/multiz5way # bash script #!/bin/sh export geneTbl="knownGene" for S in 1000 2000 5000 do echo "making upstream${S}.maf" featureBits hg38 ${geneTbl}:upstream:${S} -fa=/dev/null -bed=stdout \ | perl -wpe 's/_up[^\t]+/\t0/' | sort -k1,1 -k2,2n \ | /cluster/bin/$MACHTYPE/mafFrags hg38 multiz5way \ stdin stdout \ -orgs=/hive/data/genomes/hg38/bed/multiz5way/species.list \ | gzip -c > upstream${S}.${geneTbl}.maf.gz echo "done upstream${S}.${geneTbl}.maf.gz" done # about 20 minutes md5sum *.maf.gz *.nh upstream*.gz README.txt >> md5sum.txt # some other symlinks were already made above # obtain the README.txt from cavPor3/multiz5way and update for this # situation ln -s `pwd`/upstream*.gz `pwd`/README.txt \ /usr/local/apache/htdocs-hgdownload/goldenPath/hg38/multiz5way ############################################################################# # hgPal downloads (TBD - 2016-05-11 - Hiram) # FASTA from 5-way for knownGene, refGene and knownCanonical ssh hgwdev screen -S hg38HgPal mkdir /hive/data/genomes/hg38/bed/multiz5way/pal cd /hive/data/genomes/hg38/bed/multiz5way/pal cat ../species.list | tr '[ ]' '[\n]' > order.list # this for loop takes about 2.5 hours on this large count contig assembly export mz=multiz5way export gp=xenoRefGene export db=hg38 export I=0 export D=0 mkdir exonAA exonNuc for C in `sort -nk2 ../../../chrom.sizes | cut -f1` do I=`echo $I | awk '{print $1+1}'` D=`echo $D | awk '{print $1+1}'` dNum=`echo $D | awk '{printf "%03d", int($1/1000)}'` mkdir -p exonNuc/${dNum} > /dev/null mkdir -p exonAA/${dNum} > /dev/null echo "mafGene -chrom=$C -exons -noTrans $db $mz $gp order.list stdout | gzip -c > exonNuc/${dNum}/$C.exonNuc.fa.gz &" echo "mafGene -chrom=$C -exons $db $mz $gp order.list stdout | gzip -c > exonAA/${dNum}/$C.exonAA.fa.gz &" if [ $I -gt 16 ]; then echo "date" echo "wait" I=0 fi done > $gp.jobs echo "date" >> $gp.jobs echo "wait" >> $gp.jobs time sh -x ./$gp.jobs > $gp.jobs.log 2>&1 & # real 175m50.376s export mz=multiz5way export gp=xenoRefGene time find ./exonAA -type f | grep exonAA.fa.gz | xargs zcat \ | gzip -c > $gp.$mz.exonAA.fa.gz # real 10m29.500s time find ./exonNuc -type f | grep exonNuc.fa.gz | xargs zcat \ | gzip -c > $gp.$mz.exonNuc.fa.gz # real 15m9.974s # -rw-rw-r-- 1 611281544 Apr 15 20:37 xenoRefGene.multiz5way.exonAA.fa.gz # -rw-rw-r-- 1 955571425 Apr 15 21:06 xenoRefGene.multiz5way.exonNuc.fa.gz export mz=multiz5way export gp=xenoRefGene export db=hg38 export pd=/usr/local/apache/htdocs-hgdownload/goldenPath/$db/$mz/alignments mkdir -p $pd md5sum *.fa.gz > md5sum.txt ln -s `pwd`/$gp.$mz.exonAA.fa.gz $pd/$gp.exonAA.fa.gz ln -s `pwd`/$gp.$mz.exonNuc.fa.gz $pd/$gp.exonNuc.fa.gz ln -s `pwd`/md5sum.txt $pd/ rm -rf exonAA exonNuc ############################################################################# # wiki page for 5-way (DONE - 2017-12-18 - Hiram) mkdir /hive/users/hiram/bigWays/hg38.5way cd /hive/users/hiram/bigWays echo "hg38" > hg38.5way/ordered.list awk '{print $1}' /hive/data/genomes/hg38/bed/multiz5way/5way.distances.txt \ >> hg38.5way/ordered.list # sizeStats.sh catches up the cached measurements required for data # in the tables. They are usually already mostly done, only new # assemblies will have updates. ./sizeStats.sh hg38.5way/ordered.list # dbDb.sh constructs hg38.5way/Hg38_5-way_conservation_alignment.html # may need to add new assembly references to srcReference.list and # urlReference.list ./dbDb.sh hg38 5way # sizeStats.pl constructs hg38.5way/Hg38_5-way_Genome_size_statistics.html # this requires entries in coverage.list for new sequences ./sizeStats.pl hg38 5way # defCheck.pl constructs Hg38_5-way_conservation_lastz_parameters.html ./defCheck.pl hg38 5way # this constructs the html pages in hg38.5way/: # -rw-rw-r-- 3270 Dec 18 13:57 Hg38_5-way_conservation_alignment.html # -rw-rw-r-- 4831 Dec 18 13:57 Hg38_5-way_Genome_size_statistics.html # -rw-rw-r-- 3289 Dec 18 13:57 Hg38_5-way_conservation_lastz_parameters.html # add those pages to the genomewiki. Their page names are the # names of the .html files without the .html: # Hg38_5-way_conservation_alignment # Hg38_5-way_Genome_size_statistics # Hg38_5-way_conservation_lastz_parameters # when you view the first one you enter, it will have links to the # missing two. ############################################################################