############################################################################# ## 7way Multiz (DONE - 2019-10-15 - Hiram) ssh hgwdev mkdir /hive/data/genomes/regenRn1/bed/multiz7way cd /hive/data/genomes/regenRn1/bed/multiz7way # from the 218-way in the source tree, select out the 6 used here: /cluster/bin/phast/tree_doctor \ --prune-all-but rn6,hg38,oryCun2,mm10,canFam3,macFas5,susScr3 \ /cluster/home/hiram/kent/src/hg/utils/phyloTrees/218way.nh \ | sed -e 's/rn6/regenRn1/; s/susScr3/susScr11/;' > regenRn1.7way.nh.0 cat regenRn1.7way.nh.0 # (((hg38:0.035974,macFas5:0.043601):0.109934, # ((mm10:0.089509,regenRn1:0.096589):0.251661, # oryCun2:0.215690):0.015313):0.020593, # (susScr11:0.140000,canFam3:0.133030):0.032898); # using TreeGraph2 tree editor on the Mac, rearrange to get regenRn1 # at the top: # what that looks like: ~/kent/src/hg/utils/phyloTrees/asciiTree.pl regenRn1.7way.nh | sed -e 's/^/# /;' # ((((regenRn1:0.096589, # mm10:0.089509):0.251661, # oryCun2:0.21569):0.015313, # (hg38:0.035974, # macFas5:0.043601):0.109934):0.020593, # (susScr11:0.14, # canFam3:0.13303):0.032898); # extract species list from that .nh file sed 's/[a-z][a-z]*_//g; s/:[0-9\.][0-9\.]*//g; s/;//; /^ *$/d' \ regenRn1.7way.nh | xargs echo | sed 's/ //g; s/,/ /g' \ | sed 's/[()]//g; s/,/ /g' | tr '[ ]' '[\n]' > species.list.txt # construct db to name translation list: cat species.list.txt | while read DB do hgsql -N -e "select name,organism from dbDb where name=\"${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`" regenRn1.7way.nh | sed -e 's/00*)/)/g; s/00*,/,/g' \ | $HOME/kent/src/hg/utils/phyloTrees/asciiTree.pl /dev/stdin \ > regenRn1.7way.commonNames.nh cat regenRn1.7way.commonNames.nh | sed -e 's/^/# /;' # ((((Rat:0.096589, # Mouse:0.089509):0.251661, # Rabbit:0.21569):0.015313, # (Human:0.035974, # Crab_eating_macaque:0.043601):0.109934):0.020593, # (Pig:0.14, # Dog:0.13303):0.032898); # 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/regenRn1_7way.png ~/kent/src/hg/utils/phyloTrees/asciiTree.pl regenRn1.7way.nh > t.nh ~/kent/src/hg/utils/phyloTrees/scientificNames.sh t.nh \ | $HOME/kent/src/hg/utils/phyloTrees/asciiTree.pl /dev/stdin \ > regenRn1.7way.scientificNames.nh cat regenRn1.7way.scientificNames.nh | sed -e 's/^/# /;' # ((((Rattus_norvegicus:0.096589, # Mus_musculus:0.089509):0.251661, # Oryctolagus_cuniculus:0.21569):0.015313, # (Homo_sapiens:0.035974, # Macaca_fascicularis:0.043601):0.109934):0.020593, # (Sus_scrofa:0.14, # Canis_lupus_familiaris:0.13303):0.032898); /cluster/bin/phast/all_dists regenRn1.7way.nh | grep regenRn1 \ | sed -e "s/regenRn1.//" | sort -k2n > 7way.distances.txt # Use this output to create the table below cat 7way.distances.txt | sed -e 's/^/# /;' # mm10 0.186098 # hg38 0.509471 # macFas5 0.517098 # canFam3 0.550084 # susScr11 0.557054 # oryCun2 0.563940 printf '#!/usr/bin/env perl use strict; use warnings; open (FH, "<7way.distances.txt") or die "can not read 7way.distances.txt"; my $count = 0; while (my $line = ) { chomp $line; my ($D, $dist) = split('"'"'\\s+'"'"', $line); my $chain = "chain" . ucfirst($D); my $B="/hive/data/genomes/regenRn1/bed/lastz.$D/fb.regenRn1." . $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.regenRn1/fb.${D}.chainRegenRn1Link.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, $D; } close (FH); ' > sizeStats.pl chmod +x ./sizeStats.pl ./sizeStats.pl # If you can fill in all the numbers in this table, you are ready for # the multiple alignment procedure # featureBits chainLink measures # chainLink # N distance on regenRn1 on other other species ### type of chain: 'chain' # 01 0.1861 (% 71.606) (% 64.046) - Mouse mm10 # 02 0.5095 (% 36.240) (% 27.876) - Human hg38 # 03 0.5171 (% 35.731) (% 28.944) - Crab-eating macaque macFas5 # 04 0.5501 (% 29.714) (% 28.119) - Dog canFam3 # 05 0.5571 (% 28.152) (% 25.725) - Pig susScr11 # 06 0.5639 (% 25.702) (% 22.825) - Rabbit oryCun2 ### type of chain: 'chainSyn' # 01 0.1861 (% 68.555) (% 59.983) - Mouse mm10 # 02 0.5095 (% 34.525) (% 26.141) - Human hg38 # 03 0.5171 (% 33.940) (% 27.521) - Crab-eating macaque macFas5 # 04 0.5501 (% 28.239) (% 26.845) - Dog canFam3 # 05 0.5571 (% 26.551) (% 24.477) - Pig susScr11 # 06 0.5639 (% 23.625) (% 20.655) - Rabbit oryCun2 ### type of chain: 'chainRBest.' # 01 0.1861 (% 68.579) (% 59.001) - Mouse mm10 # 02 0.5095 (% 34.786) (% 25.599) - Human hg38 # 03 0.5171 (% 34.286) (% 27.870) - Crab-eating macaque macFas5 # 04 0.5501 (% 28.564) (% 27.216) - Dog canFam3 # 05 0.5571 (% 26.920) (% 24.832) - Pig susScr11 # 06 0.5639 (% 24.463) (% 21.426) - Rabbit oryCun2 # None of this concern for distances matters in building the first step, the # maf files. The distances will be better calibrated later. # create species list and stripped down tree for autoMZ sed 's/[a-z][a-z]*_//g; s/:[0-9\.][0-9\.]*//g; s/;//; /^ *$/d' \ regenRn1.7way.nh | xargs echo | sed 's/ //g; s/,/ /g' > tree.nh sed 's/[()]//g; s/,/ /g' tree.nh > species.list # regenRn1 mm10 oryCun2 hg38 macFas5 susScr11 canFam3 # survey N60 for each for db in `cat species.list` do n50.pl /hive/data/genomes/$db/chrom.sizes 2>&1 | head -4 | grep -v N50 done # reading: /hive/data/genomes/regenRn1/chrom.sizes # contig count: 23, total size: 2395213054, one half size: 1197606527 1145314132 7 chr8 119798696 # reading: /hive/data/genomes/mm10/chrom.sizes # contig count: 66, total size: 2730871774, one half size: 1365435887 1312176979 8 chr7 145441459 # reading: /hive/data/genomes/oryCun2/chrom.sizes # contig count: 3242, total size: 2737490501, one half size: 1368745250 1277423395 8 chr9 116251907 # reading: /hive/data/genomes/hg38/chrom.sizes # contig count: 595, total size: 3257347282, one half size: 1628673641 1547391171 8 chrX 156040895 # reading: /hive/data/genomes/macFas5/chrom.sizes # contig count: 7601, total size: 2946843737, one half size: 1473421868 1326187189 7 chr4 170955103 # reading: /hive/data/genomes/susScr11/chrom.sizes # contig count: 613, total size: 2501912388, one half size: 1250956194 1227124957 7 chr9 139512083 # reading: /hive/data/genomes/canFam3/chrom.sizes # contig count: 3268, total size: 2410976875, one half size: 1205488437 1178632958 14 chr15 64190966 # bash shell syntax here ... cd /hive/data/genomes/regenRn1/bed/multiz7way export H=/hive/data/genomes/regenRn1/bed mkdir mafLinks # these are all good assemblies, can use syntenic net: for G in mm10 oryCun2 hg38 macFas5 susScr11 canFam3 do mkdir mafLinks/$G echo ln -s ${H}/lastz.$G/axtChain/regenRn1.${G}.synNet.maf.gz ./mafLinks/$G ln -s ${H}/lastz.$G/axtChain/regenRn1.${G}.synNet.maf.gz ./mafLinks/$G done # verify the symLinks are good: ls -ogrtL mafLinks/*/* | sed -e 's/^/# /; s/-rw-rw-r-- 1//;' # 612950682 Sep 13 12:43 mafLinks/hg38/regenRn1.hg38.synNet.maf.gz # 1034934817 Sep 13 13:12 mafLinks/mm10/regenRn1.mm10.synNet.maf.gz # 416798141 Oct 11 15:15 mafLinks/oryCun2/regenRn1.oryCun2.synNet.maf.gz # 502502792 Oct 14 14:19 mafLinks/canFam3/regenRn1.canFam3.synNet.maf.gz # 471853744 Oct 14 14:21 mafLinks/susScr11/regenRn1.susScr11.synNet.maf.gz # 602122175 Oct 14 15:09 mafLinks/macFas5/regenRn1.macFas5.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/regenRn1/bed/multiz7way/mafSplit cd /hive/data/genomes/regenRn1/bed/multiz7way/mafSplit time for D in `sed -e "s/regenRn1 //" ../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/${D}/*.maf.gz cd .. done # real 2m5.470s # construct a list of all possible maf file names. # they do not all exist in each of the species directories find . -type f | wc -l # 126 find . -type f | grep ".maf$" | xargs -L 1 basename | sort -u > maf.list wc -l maf.list # 21 maf.list mkdir /hive/data/genomes/regenRn1/bed/multiz7way/splitRun cd /hive/data/genomes/regenRn1/bed/multiz7way/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 = regenRn1 set c = $1 set result = $2 set run = `/bin/pwd` set tmp = /dev/shm/$db/multiz.$c set pairs = /hive/data/genomes/regenRn1/bed/multiz7way/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 $(file1) {check out line+ /hive/data/genomes/regenRn1/bed/multiz7way/splitRun/maf/$(root1).maf} #ENDLOOP ' > template ln -s ../../mafSplit/maf.list maf.list ssh ku cd /hive/data/genomes/regenRn1/bed/multiz7way/splitRun/run gensub2 maf.list single template jobList para create jobList para try ... check ... push ... etc... # Completed: 21 of 21 jobs # CPU time in finished jobs: 83062s 1384.37m 23.07h 0.96d 0.003 y # IO & Wait Time: 74s 1.23m 0.02h 0.00d 0.000 y # Average job time: 3959s 65.98m 1.10h 0.05d # Longest finished job: 8511s 141.85m 2.36h 0.10d # Submission to last job: 8518s 141.97m 2.37h 0.10d # combine into one file (the 1>&2 redirect sends the echo to stderr) cd /hive/data/genomes/regenRn1/bed/multiz7way head -1 splitRun/maf/217.maf > multiz7way.maf time for F in splitRun/maf/*.maf do echo "${F}" 1>&2 egrep -v "^#" ${F} done >> multiz7way.maf # real 1m12.634s tail -1 splitRun/maf/217.maf >> multiz7way.maf # -rw-rw-r-- 1 9942794566 Oct 16 09:24 multiz7way.maf # Load into database ssh hgwdev cd /hive/data/genomes/regenRn1/bed/multiz7way mkdir /gbdb/regenRn1/multiz7way ln -s `pwd`/multiz7way.maf /gbdb/regenRn1/multiz7way cd /dev/shm time hgLoadMaf regenRn1 multiz7way # Loaded 8770521 mafs in 1 files from /gbdb/regenRn1/multiz7way # real 1m38.632s time hgLoadMafSummary -verbose=2 -minSize=30000 \ -mergeGap=1500 -maxSize=200000 regenRn1 multiz7waySummary \ /gbdb/regenRn1/multiz7way/multiz7way.maf # Created 1206135 summary blocks from 28567597 components and 8770521 mafs from /gbdb/regenRn1/multiz7way/multiz7way.maf # real 2m2.016s # -rw-rw-r-- 1 439436348 Oct 16 09:26 multiz7way.tab # -rw-rw-r-- 1 55980922 Oct 16 09:39 multiz7waySummary.tab wc -l multiz7way*.tab # 8770521 multiz7way.tab # 1206135 multiz7waySummary.tab rm multiz7way*.tab ############################################################################## # GAP ANNOTATE multiz7way MAF AND LOAD TABLES (DONE - 2017-12-13 - Hiram) # mafAddIRows has to be run on single chromosome maf files, it does not # function correctly when more than one reference sequence # are in a single file. Need to split of the maf file into individual # maf files mkdir -p /hive/data/genomes/regenRn1/bed/multiz7way/anno/mafSplit cd /hive/data/genomes/regenRn1/bed/multiz7way/anno/mafSplit time mafSplit -outDirDepth=2 -byTarget -useFullSequenceName \ /dev/null . ../../multiz7way.maf # real 1m54.035s find . -type f | wc -l # 22 # check for N.bed files everywhere: cd /hive/data/genomes/regenRn1/bed/multiz7way/anno for DB in `cat ../species.list` do if [ ! -s /hive/data/genomes/${DB}/${DB}.N.bed ]; then echo "MISS: ${DB}" # cd /hive/data/genomes/${DB} # twoBitInfo -nBed ${DB}.2bit ${DB}.N.bed else echo " OK: ${DB}" fi done cd /hive/data/genomes/regenRn1/bed/multiz7way/anno for DB in `cat ../species.list` do echo "${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 ${DB}.len >> sizes done # make sure they all are successful symLinks: ls -ogrtL screen -S gapAnno # use a screen to control this longish job ssh ku cd /hive/data/genomes/regenRn1/bed/multiz7way/anno mkdir result find ./mafSplit -type d | sed -e 's#./mafSplit/##' | while read D do echo mkdir -p result/${D} mkdir -p result/${D} done printf '#LOOP mafAddIRows -nBeds=nBeds mafSplit/$(path1) /hive/data/genomes/regenRn1/regenRn1.2bit {check out exists+ result/$(path1)} #ENDLOOP ' > template find ./mafSplit -type f | sed -e 's#^./mafSplit/##' > maf.list gensub2 maf.list single template jobList # limit jobs on a node with the ram=32g requirement because they go fast para -ram=32g create jobList para try ... check ... push ... # Completed: 22 of 22 jobs # CPU time in finished jobs: 252s 4.20m 0.07h 0.00d 0.000 y # IO & Wait Time: 78s 1.30m 0.02h 0.00d 0.000 y # Average job time: 15s 0.25m 0.00h 0.00d # Longest finished job: 31s 0.52m 0.01h 0.00d # Submission to last job: 98s 1.63m 0.03h 0.00d # verify all result files have some content, look for 0 size files: find ./result -type f -size 0 # should see none # or in this manner: find ./result -type f | xargs ls -og | sort -k3nr | tail # combine into one file (the 1>&2 redirect sends the echo to stderr) head -q -n 1 result/0/2/chr1.maf > regenRn1.7way.maf time find ./result -type f | while read F do echo "${F}" 1>&2 grep -h -v "^#" ${F} done >> regenRn1.7way.maf # real 1m22.625s # these maf files do not have the end marker, this does nothing: # tail -q -n 1 result/0/0/NW_007781932v1.maf >> regenRn1.7way.maf # How about an official end marker: echo "##eof maf" >> regenRn1.7way.maf ls -og # -rw-rw-r-- 1 12551361334 Oct 16 10:13 regenRn1.7way.maf du -hsc regenRn1.7way.maf ../*.maf # 12G regenRn1.7way.maf # 9.3G ../multiz7way.maf # construct symlinks to get the individual maf files into gbdb: rm /gbdb/regenRn1/multiz7way/multiz7way.maf # remove previous results ln -s `pwd`/regenRn1.7way.maf /gbdb/regenRn1/multiz7way/multiz7way.maf # Load into database cd /dev/shm time hgLoadMaf -pathPrefix=/gbdb/regenRn1/multiz7way regenRn1 multiz7way # Loaded 9319931 mafs in 1 files from /gbdb/regenRn1/multiz7way # real 2m5.578s time hgLoadMafSummary -verbose=2 -minSize=30000 \ -mergeGap=1500 -maxSize=200000 regenRn1 multiz7waySummary \ /gbdb/regenRn1/multiz7way/multiz7way.maf # Created 1206135 summary blocks from 28567597 components and 9319931 mafs from /gbdb/regenRn1/multiz7way/multiz7way.maf # real 2m20.161s # -rw-rw-r-- 1 469026908 Oct 16 10:16 multiz7way.tab # -rw-rw-r-- 1 58393192 Oct 16 10:19 multiz7waySummary.tab rm multiz7way*.tab ###################################################################### # multiz7way MAF FRAMES (TBD - 2016-06-06 - Hiram) ssh hgwdev mkdir /hive/data/genomes/regenRn1/bed/multiz7way/frames cd /hive/data/genomes/regenRn1/bed/multiz7way/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 chmod +x ./showGenes.csh time ./showGenes.csh # regenRn1: mgcGenes: 6243, refGene: 16621, xenoRefGene: 195943, Mrnas: 1252257 # mm10: ensGene: 103734, knownGene: 142446, mgcGenes: 27846, ncbiRefSeq: 106520, refGene: 44299, xenoRefGene: 187341, Mrnas: 5492915 # oryCun2: ensGene: 24964, refGene: 1746, xenoRefGene: 335024, Mrnas: 40860 # hg38: ensGene: 208239, knownGene: 247541, mgcGenes: 36653, ncbiRefSeq: 167469, refGene: 82864, xenoRefGene: 197482, Mrnas: 11695943 # macFas5: ensGene: 54367, ncbiRefSeq: 76183, refGene: 2160, xenoRefGene: 328224, Mrnas: 179478 # susScr11: ensGene: 49448, ncbiRefSeq: 77695, refGene: 4599, xenoRefGene: 218711, Mrnas: 1722885 # canFam3: ensGene: 39074, ncbiRefSeq: 82536, refGene: 2303, xenoRefGene: 270976, Mrnas: 388203 real 1m7.259s # from that summary, use these gene sets: # knownGene - hg38 mm10 # refGene - regenRn1 # ensGene - oryCun2 # ncbiRefSeq - macFas5 canFam3 susScr11 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: 22100 failed: 0 # mm10: checked: 22026 failed: 0 # 2. ensGene: oryCun2 for DB in oryCun2 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 # oryCun2: checked: 19165 failed: 0 # 3. refGene: regenRn1 for DB in regenRn1 do hgsql -N -e "select name,chrom,strand,txStart,txEnd,cdsStart,cdsEnd,exonCount,exonStarts,exonEnds from refGene" ${DB} \ | genePredSingleCover stdin stdout | gzip -2c \ > /dev/shm/${DB}.tmp.gz mv /dev/shm/${DB}.tmp.gz genes/$DB.gp.gz printf "# ${DB}: " genePredCheck -db=${DB} genes/${DB}.gp.gz done # regenRn1: checked: 14952 failed: 0 # 4. ncbiRefSeq - macFas5 canFam3 susScr11 for DB in macFas5 canFam3 susScr11 do hgsql -N -e "select name,chrom,strand,txStart,txEnd,cdsStart,cdsEnd,exonCount,exonStarts,exonEnds from ncbiRefSeq" ${DB} \ | genePredSingleCover stdin stdout | gzip -2c \ > /dev/shm/${DB}.tmp.gz mv /dev/shm/${DB}.tmp.gz genes/$DB.gp.gz printf "# ${DB}: " genePredCheck -db=${DB} genes/${DB}.gp.gz done # macFas5: checked: 20552 failed: 0 # canFam3: checked: 19805 failed: 0 # susScr11: checked: 20615 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/canFam3.gp.gz: 19803 # genes/hg38.gp.gz: 22082 # genes/macFas5.gp.gz: 20521 # genes/mm10.gp.gz: 22026 # genes/oryCun2.gp.gz: 19165 # genes/regenRn1.gp.gz: 14897 # genes/susScr11.gp.gz: 20614 time (cat ../anno/regenRn1.7way.maf \ | genePredToMafFrames regenRn1 stdin stdout \ `cat ../species.list.txt | xargs echo \ | sed -e "s#\([a-zA-Z0-9]*\)#\1 genes/\1.gp.gz#g;"` \ | gzip > multiz7wayFrames.bed.gz) # real 1m55.421s # verify there are frames on everything, should be 4 species: zcat multiz7wayFrames.bed.gz | awk '{print $4}' | sort | uniq -c \ | sed -e 's/^/# /;' # 223345 canFam3 # 204028 hg38 # 218850 macFas5 # 191961 mm10 # 184406 oryCun2 # 137219 regenRn1 # 226281 susScr11 # load the resulting file ssh hgwdev cd /hive/data/genomes/regenRn1/bed/multiz7way/frames time hgLoadMafFrames regenRn1 multiz7wayFrames multiz7wayFrames.bed.gz # real 0m10.853s time featureBits -countGaps regenRn1 multiz7wayFrames # 34777903 bases of 2395213054 (1.452%) in intersection # real 0m8.330s # enable the trackDb entries: # frames multiz7wayFrames # irows on # appears to work OK ######################################################################### # Phylogenetic tree from 7way (DONE - 2017-12-17 - Hiram) mkdir /hive/data/genomes/regenRn1/bed/multiz7way/4d cd /hive/data/genomes/regenRn1/bed/multiz7way/4d # using the refGene hgsql -N -e "select name,chrom,strand,txStart,txEnd,cdsStart,cdsEnd,exonCount,exonStarts,exonEnds from refGene" regenRn1 \ | genePredSingleCover stdin stdout > regenRn1.xenoRefGeneNR.gp genePredCheck -db=regenRn1 regenRn1.refGeneNR.gp # checked: 14952 failed: 0 # the annotated maf is: og ../anno/regenRn1.7way.maf # -rw-rw-r-- 1 12551361334 Oct 16 10:13 ../anno/regenRn1.7way.maf mkdir annoSplit cd annoSplit time mafSplit -verbose=2 -outDirDepth=2 -byTarget -useFullSequenceName \ /dev/null . ../../anno/regenRn1.7way.maf # real 2m37.599s find . -type f | wc -l # 22 ssh ku mkdir /hive/data/genomes/regenRn1/bed/multiz7way/4d/run cd /hive/data/genomes/regenRn1/bed/multiz7way/4d/run mkdir ../mfa # newer versions of msa_view have a slightly different operation # the sed of the gp file inserts the reference species in the chr name cat << '_EOF_' > 4d.csh #!/bin/csh -fex set PHASTBIN = /cluster/bin/phast.build/cornellCVS/phast.2010-12-30/bin set GP = regenRn1.refGeneNR.gp set r = "/hive/data/genomes/regenRn1/bed/multiz7way" 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$c\t/\tregenRn1.$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 # 22 maf.list printf '#LOOP 4d.csh $(root1) annoSplit/$(dir1)/$(file1) mfa/$(dir1) {check out line+ ../mfa/$(dir1)/$(root1).mfa} #ENDLOOP ' > template mkdir ../mfa gensub2 maf.list single template jobList para create jobList para try ... check para time # Completed: 21 of 22 jobs # Crashed: 1 jobs # CPU time in finished jobs: 549s 9.15m 0.15h 0.01d 0.000 y # IO & Wait Time: 58s 0.97m 0.02h 0.00d 0.000 y # Average job time: 29s 0.48m 0.01h 0.00d # Longest finished job: 58s 0.97m 0.02h 0.00d # Submission to last job: 1187s 19.78m 0.33h 0.01d # 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/regenRn1/bed/multiz7way/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, it isn't safe, it # outputs files that are larger than size 1: ### XXX 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 $NF}' \ > empty.list cat empty.list | xargs rm -f # see what is left: ls -ogrt mfa/*/*/*.mfa | sort -k3nr | wc # 21 147 1082 # want comma-less species.list time /cluster/bin/phast.build/cornellCVS/phast.2010-12-30/bin/msa_view \ --aggregate "`cat ../species.list`" mfa/*/*/*.mfa | sed s/"> "/">"/ \ > 4d.all.mfa # real 0m1.239s # check they are all in there: grep "^>" 4d.all.mfa | sed -e 's/^/# /;' # >regenRn1 # >mm10 # >oryCun2 # >hg38 # >macFas5 # >susScr11 # >canFam3 sed 's/[a-z][a-z]*_//g; s/:[0-9\.][0-9\.]*//g; s/;//; /^ *$/d' \ ../regenRn1.7way.nh | xargs echo | sed -e 's/ //g' > tree_commas.nh # tree_commas.nh looks like: # ((((regenRn1,mm10),oryCun2),(hg38,macFas5)),(susScr11,canFam3)) # 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 0m3.050s mv phyloFit.mod all.mod grep TREE all.mod # TREE: ((((regenRn1:0.0862581,mm10:0.081071):0.240213, # oryCun2:0.1895):0.0217336, # (hg38:0.0298154,macFas5:0.0357728):0.0939174):0.0214404, # (susScr11:0.151716,canFam3:0.14205):0.0214404); # compare these calculated lengths to the tree extracted from 191way: grep TREE all.mod | sed -e 's/TREE: //' \ | /cluster/bin/phast/all_dists /dev/stdin | grep regenRn1 \ | sed -e "s/regenRn1.//;" | sort > new.dists /cluster/bin/phast/all_dists ../regenRn1.7way.nh | grep regenRn1 \ | sed -e "s/regenRn1.//;" | sort > old.dists # printing out the 'new', the 'old' the 'difference' and percent difference join new.dists old.dists | awk '{ printf "#\t%s\t%8.5f\t%8.5f\t%8.5f\t%8.5f\n", $1, $2, $3, $2-$3, 100*($2-$3)/$3 }' \ | sort -k3n # mm10 0.16733 0.18610 -0.01877 -10.08555 # hg38 0.47194 0.50947 -0.03753 -7.36705 # macFas5 0.47790 0.51710 -0.03920 -7.58135 # oryCun2 0.51597 0.56394 -0.04797 -8.50605 # canFam3 0.53314 0.55008 -0.01695 -3.08098 # susScr11 0.54280 0.55705 -0.01425 -2.55846 ######################################################################### # phastCons 7way (TBD - 2016-06-06 - Hiram) # split 7way mafs into 10M chunks and generate sufficient statistics # files for # phastCons ssh ku mkdir -p /hive/data/genomes/regenRn1/bed/multiz7way/cons/SS cd /hive/data/genomes/regenRn1/bed/multiz7way/cons/SS mkdir result done printf '#!/bin/csh -ef set d = $1 set c = $2 set doneDir = done/$d set MAF = /hive/data/genomes/regenRn1/bed/multiz7way/anno/result/$d/$c.maf set WINDOWS = /hive/data/genomes/regenRn1/bed/multiz7way/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 # 22 maf.list ssh ku cd /hive/data/genomes/regenRn1/bed/multiz7way/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: 22 of 22 jobs # CPU time in finished jobs: 817s 13.62m 0.23h 0.01d 0.000 y # IO & Wait Time: 223s 3.71m 0.06h 0.00d 0.000 y # Average job time: 47s 0.79m 0.01h 0.00d # Longest finished job: 91s 1.52m 0.03h 0.00d # Submission to last job: 269s 4.48m 0.07h 0.00d find ./result -type f | wc -l # 252 # 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/regenRn1/bed/multiz7way/cons/run.cons cd /hive/data/genomes/regenRn1/bed/multiz7way/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/regenRn1/bed/multiz7way/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)/$(root1).pp} #ENDLOOP ' > template find ../SS/result -type f | sed -e "s#../SS/result/##" > ss.list wc -l ss.list # 252 ss.list # Create parasol batch and run it # run for all species cd /hive/data/genomes/regenRn1/bed/multiz7way/cons mkdir -p all cd all # Using the .mod tree cp -p ../../4d/all.mod ./all.mod gensub2 ../run.cons/ss.list single ../run.cons/template jobList para -ram=32g create jobList para try ... check ... para push # Completed: 252 of 252 jobs # CPU time in finished jobs: 3353s 55.88m 0.93h 0.04d 0.000 y # IO & Wait Time: 1653s 27.55m 0.46h 0.02d 0.000 y # Average job time: 20s 0.33m 0.01h 0.00d # Longest finished job: 26s 0.43m 0.01h 0.00d # Submission to last job: 168s 2.80m 0.05h 0.00d # create Most Conserved track cd /hive/data/genomes/regenRn1/bed/multiz7way/cons/all time cut -f1 ../../../../chrom.sizes | while read C do ls -d bed/?/?/${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 0m11.331s time /cluster/bin/scripts/lodToBedScore tmpMostConserved.bed \ > mostConserved.bed # real 0m8.310s # -rw-rw-r-- 1 36177141 Oct 16 14:23 mostConserved.bed # load into database ssh hgwdev cd /hive/data/genomes/regenRn1/bed/multiz7way/cons/all time hgLoadBed regenRn1 phastConsElements7way mostConserved.bed # Read 1039897 elements of size 5 from mostConserved.bed # real 0m5.439s # on human we often try for 6% 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 46 --target-coverage 0.3 time featureBits regenRn1 -enrichment refGene:cds phastConsElements7way # refGene:cds 1.024%, phastConsElements7way 4.734%, both 0.784%, # cover 76.58%, enrich 16.18x # real 0m8.156s # Create merged posterier probability file and wiggle track data files cd /hive/data/genomes/regenRn1/bed/multiz7way/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/phastCons7way.wigFix.gz) # about 15 to 20 minutes # -rw-rw-r-- 1 2128227895 Oct 16 14:40 phastCons7way.wigFix.gz # check integrity of data with wigToBigWig time (zcat downloads/phastCons7way.wigFix.gz \ | wigToBigWig -verbose=2 stdin /hive/data/genomes/regenRn1/chrom.sizes \ phastCons7way.bw) > bigWig.log 2>&1 egrep "real|VmPeak" bigWig.log # pid=129403: VmPeak: 18745464 kB # real 18m20.780s bigWigInfo phastCons7way.bw | sed -e 's/^/# /;' # version: 4 # isCompressed: yes # isSwapped: 0 # primaryDataSize: 3,231,215,692 # primaryIndexSize: 62,912,880 # zoomLevels: 10 # chromCount: 22 # basesCovered: 1,694,662,819 # mean: 0.150180 # min: 0.000000 # max: 1.000000 # std: 0.254162 # encode those files into wiggle data time (zcat downloads/phastCons7way.wigFix.gz \ | wigEncode stdin phastCons7way.wig phastCons7way.wib) # Converted stdin, upper limit 1.00, lower limit 0.00 # real 7m14.664s du -hsc *.wi? # 1.6G phastCons7way.wib # 187M phastCons7way.wig # Load gbdb and database with wiggle. ln -s `pwd`/phastCons7way.wib /gbdb/regenRn1/multiz7way/phastCons7way.wib time hgLoadWiggle -pathPrefix=/gbdb/regenRn1/multiz7way \ regenRn1 phastCons7way phastCons7way.wig # real 0m12.453s # use to set trackDb.ra entries for wiggle min and max # and verify table is loaded correctly wigTableStats.sh regenRn1 phastCons7way # db.table min max mean count sumData # regenRn1.phastCons7way 0 1 0.15018 1694662819 2.54505e+08 # stdDev viewLimits # 0.254162 viewLimits=0:1 # Create histogram to get an overview of all the data # Do not mix stderr with the output, it gets confused time hgWiggle -doHistogram -db=regenRn1 \ -hBinSize=0.001 -hBinCount=1000 -hMinVal=0.0 -verbose=2 \ phastCons7way > regenRn1.phastCons7way.histogram.data 2> t.err # real 1m15.914s # the Y axis range: grep -v "^#" regenRn1.phastCons7way.histogram.data | awk '{print $5}' | ave stdin # Q1 0.000107 # median 0.000218 # Q3 0.000547 # average 0.001000 # min 0.000012 # max 0.240878 # count 1000 # total 1.000008 # standard deviation 0.007900 # create plot of histogram: # updated for new gnuplot on hgwdev 2018-11-26 (can't get font to change) printf 'set terminal pngcairo size 1000,600 background "#000000" font "/usr/share/fonts/default/Type1/n022004l.pfb" set output "regenRn1.phastCons7way.histo.png" set size 1.0, 1.0 set style line 1 lt 2 lc rgb "#ff88ff" lw 2 set style line 2 lt 2 lc rgb "#66ff66" lw 2 set style line 3 lt 2 lc rgb "#ffff00" lw 2 set style line 4 lt 2 lc rgb "#ffffff" lw 2 set border lc rgb "#ffff00" set key left box ls 3 set key tc variable set grid noxtics set y2tics set grid ytics ls 4 set title " Rat/regenRn1 Histogram phastCons7way track" \ tc rgb "#ffffff" set xlabel " phastCons7way score" tc rgb "#ffffff" set ylabel " Relative Frequency" tc rgb "#ff88ff" set y2label " Cumulative Relative Frequency (CRF)" tc rgb "#66ff66" set y2range [0:1] set yrange [0:0.25] plot "regenRn1.phastCons7way.histogram.data" using 2:5 title " RelFreq" with impulses ls 1, \ "regenRn1.phastCons7way.histogram.data" using 2:7 axes x1y2 title " CRF" with lines ls 2 ' | gnuplot display regenRn1.phastCons7way.histo.png & ######################################################################### # phyloP for 7way (TBD - 2016-06-09,11 - Hiram) # run phyloP with score=LRT ssh ku mkdir /cluster/data/regenRn1/bed/multiz7way/consPhyloP cd /cluster/data/regenRn1/bed/multiz7way/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}' # 0.584 /cluster/bin/phast.build/cornellCVS/phast.2010-12-30/bin/modFreqs \ ../../4d/all.mod 0.584 > all.mod # verify, the BACKGROUND should now be paired up: grep BACK all.mod # BACKGROUND: 0.208000 0.292000 0.292000 0.208000 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/regenRn1/bed/multiz7way/consPhyloP set tmp = $cons/tmp/$grp/$f /bin/rm -fr $tmp /bin/mkdir -p $tmp set ssSrc = "/hive/data/genomes/regenRn1/bed/multiz7way/cons/SS/result/$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 # 252 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/regenRn1/bed/multiz7way/consPhyloP/all cd /hive/data/genomes/regenRn1/bed/multiz7way/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=63 push para time > run.time # Completed: 252 of 252 jobs # CPU time in finished jobs: 4205s 70.08m 1.17h 0.05d 0.000 y # IO & Wait Time: 1675s 27.92m 0.47h 0.02d 0.000 y # Average job time: 23s 0.39m 0.01h 0.00d # Longest finished job: 31s 0.52m 0.01h 0.00d # Submission to last job: 240s 4.00m 0.07h 0.00d 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/phyloP7way.wigFix.gz) # real 14m25.749s # check integrity of data with wigToBigWig time (zcat downloads/phyloP7way.wigFix.gz \ | wigToBigWig -verbose=2 stdin /hive/data/genomes/regenRn1/chrom.sizes \ phyloP7way.bw) > bigWig.log 2>&1 egrep "real|VmPeak" bigWig.log # pid=45264: VmPeak: 18745464 kB # real 17m48.730s bigWigInfo phyloP7way.bw | sed -e 's/^/# /;' # version: 4 # isCompressed: yes # isSwapped: 0 # primaryDataSize: 2,570,662,548 # primaryIndexSize: 62,912,880 # zoomLevels: 10 # chromCount: 22 # basesCovered: 1,694,662,819 # mean: 0.082710 # min: -4.373000 # max: 0.967000 # std: 0.593297 # encode those files into wiggle data time (zcat downloads/phyloP7way.wigFix.gz \ | wigEncode stdin phyloP7way.wig phyloP7way.wib) # Converted stdin, upper limit 0.97, lower limit -4.37 # real 7m29.488s du -hsc *.wi? # 1.6G phyloP7way.wib # 189M phyloP7way.wig # Load gbdb and database with wiggle. ln -s `pwd`/phyloP7way.wib /gbdb/regenRn1/multiz7way/phyloP7way.wib time hgLoadWiggle -pathPrefix=/gbdb/regenRn1/multiz7way regenRn1 \ phyloP7way phyloP7way.wig # real 0m12.985s # use to set trackDb.ra entries for wiggle min and max # and verify table is loaded correctly wigTableStats.sh regenRn1 phyloP7way # db.table min max mean count sumData # regenRn1.phyloP7way -4.373 0.967 0.0827105 1694662819 1.40166e+08 # stdDev viewLimits # 0.593297 viewLimits=-2.88378:0.967 # that range is: 4.373+0.967 = 5.340 for hBinSize=0.00534 # Create histogram to get an overview of all the data # do NOT mix up stderr with the output, it interferes with the output time (hgWiggle -doHistogram \ -hBinSize=0.00534 -hBinCount=1000 -hMinVal=-4.373 -verbose=2 \ -db=regenRn1 phyloP7way) > regenRn1.phyloP7way.histogram.data \ 2> t.err # real 1m15.999s # find the Y range for the 2:6 graph grep "^[0-9]" regenRn1.phyloP7way.histogram.data | ave -col=5 stdin \ | sed -e 's/^/# /;' # Q1 0.000003 # median 0.000044 # Q3 0.000888 # average 0.001037 # min 0.000000 # max 0.059670 # count 964 # total 0.999981 # standard deviation 0.003914 # find the X range for the 2:6 graph grep "^[0-9]" regenRn1.phyloP7way.histogram.data | ave -col=2 stdin \ | sed -e 's/^/# /;' # Q1 -3.088730 # median -1.801790 # Q3 -0.514850 # average -1.778824 # min -4.373000 # max 0.967000 # count 964 # total -1714.785920 # standard deviation 1.522294 # create plot of histogram: # updated for new gnuplot on hgwdev 2018-11-26 (can't get font to change) printf 'set terminal pngcairo size 1000,600 background "#000000" font "/usr/share/fonts/default/Type1/n022004l.pfb" set output "regenRn1.phyloP7way.histo.png" set size 1.0, 1.0 set style line 1 lt 2 lc rgb "#ff88ff" lw 2 set style line 2 lt 2 lc rgb "#66ff66" lw 2 set style line 3 lt 2 lc rgb "#ffff00" lw 2 set style line 4 lt 2 lc rgb "#ffffff" lw 2 set border lc rgb "#ffff00" set key left box ls 3 set key tc variable set grid noxtics set y2tics set grid ytics ls 4 set title " Rat/regenRn1 Histogram phyloP7way track" \ tc rgb "#ffffff" set xlabel " phyloP7way score" tc rgb "#ffffff" set ylabel " Relative Frequency" tc rgb "#ff88ff" set y2label " Cumulative Relative Frequency (CRF)" tc rgb "#66ff66" set y2range [0:1] set xrange [-4.4:0.97] set yrange [0:0.06] plot "regenRn1.phyloP7way.histogram.data" using 2:5 title " RelFreq" with impulses ls 1, \ "regenRn1.phyloP7way.histogram.data" using 2:7 axes x1y2 title " CRF" with lines ls 2 ' | gnuplot # set xrange [-4.4:0.97] display regenRn1.phyloP7way.histo.png & # appears to have an odd hole in the data near X=0 ? ############################################################################# # hgPal downloads (TBD - 2016-06-09,11 - Hiram) # FASTA from 7way for refGene ssh hgwdev screen -S regenRn1HgPal mkdir /hive/data/genomes/regenRn1/bed/multiz7way/pal cd /hive/data/genomes/regenRn1/bed/multiz7way/pal cat ../species.list | tr '[ ]' '[\n]' > order.list # this for loop takes about 2.6 hours on this large count contig assembly export mz=multiz7way export gp=refGene export db=regenRn1 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 0m0.680s echo "date" >> $gp.jobs echo "wait" >> $gp.jobs chmod +x $gp.jobs time (./$gp.jobs) > $gp.jobs.log 2>&1 & # real 2m39.932s export mz=multiz7way export gp=refGene time find ./exonAA -type f | grep exonAA.fa.gz | xargs zcat \ | gzip -c > $gp.$mz.exonAA.fa.gz # real 0m5.683s time find ./exonNuc -type f | grep exonNuc.fa.gz | xargs zcat \ | gzip -c > $gp.$mz.exonNuc.fa.gz # real 0m24.455s # -rw-rw-r-- 1 28108399 Oct 17 10:54 refGene.multiz7way.exonAA.fa.gz # -rw-rw-r-- 1 46239299 Oct 17 10:54 refGene.multiz7way.exonNuc.fa.gz export mz=multiz7way export gp=refGene export db=regenRn1 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 ############################################################################# XXX - ready to go - Thu Oct 17 10:57:20 PDT 2019 # construct download files for 7way (TBD - 2016-06-11 - Hiram) mkdir /usr/local/apache/htdocs-hgdownload/goldenPath/regenRn1/multiz7way mkdir /usr/local/apache/htdocs-hgdownload/goldenPath/regenRn1/phastCons7way mkdir /usr/local/apache/htdocs-hgdownload/goldenPath/regenRn1/phyloP7way mkdir /hive/data/genomes/regenRn1/bed/multiz7way/downloads cd /hive/data/genomes/regenRn1/bed/multiz7way/downloads mkdir multiz7way phastCons7way phyloP7way cd multiz7way time cp -p ../../anno/regenRn1.7way.maf . # real 0m23.199s # -rw-rw-r-- 1 12551361334 Oct 16 10:13 regenRn1.7way.maf du -hsc * # 12G regenRn1.7way.maf time gzip *.maf # real 43m18.329s # real 22m31.598s # -rw-rw-r-- 1 3005396685 Oct 16 10:13 regenRn1.7way.maf.gz du -hsc *.maf.gz ../../anno/*.maf # 2.8G regenRn1.7way.maf.gz # 12G ../../anno/regenRn1.7way.maf grep TREE ../../4d/all.mod | awk '{print $NF}' \ | ~/kent/src/hg/utils/phyloTrees/asciiTree.pl /dev/stdin \ > regenRn1.7way.nh ln -s ../../regenRn1.7way.commonNames.nh . ln -s ../../regenRn1.7way.scientificNames.nh time md5sum *.nh *.maf.gz > md5sum.txt # real 0m36.144s ln -s `pwd`/* \ /usr/local/apache/htdocs-hgdownload/goldenPath/regenRn1/multiz7way du -hsc *.maf.gz ../../anno/regenRn1.7way.maf # 3.0G regenRn1.7way.maf.gz # 13G ../../anno/regenRn1.7way.maf # obtain the README.txt from galGal7/multiz77way and update for this # situation # to get the species table list for the README: for F in `cat ../../species.list` do hgsql -N -e "select organism,scientificName,description from dbDb where name=\"$F\";" hgcentraltest done | cat Rat Rattus norvegicus May. 2019 (Regen Rn1/regenRn1) Mouse Mus musculus Dec. 2011 (GRCm38/mm10) Rabbit Oryctolagus cuniculus Apr. 2009 (Broad/oryCun2) Human Homo sapiens Dec. 2013 (GRCh38/hg38) Crab-eating macaque Macaca fascicularis Jun. 2013 (Macaca_fascicularis_5.0/macFas5) Pig Sus scrofa Feb. 2017 (Sscrofa11.1/susScr11) Dog Canis lupus familiaris Sep. 2011 (Broad CanFam3.1/canFam3) ##################################################################### cd /hive/data/genomes/regenRn1/bed/multiz7way/downloads/phastCons7way ln -s ../../cons/all/downloads/phastCons7way.wigFix.gz \ ./regenRn1.phastCons7way.wigFix.gz ln -s ../../cons/all/phastCons7way.bw ./regenRn1.phastCons7way.bw ln -s ../../cons/all/all.mod ./regenRn1.phastCons7way.mod time md5sum *.gz *.mod *.bw > md5sum.txt # real 0m15.741s # obtain the README.txt from galGal6/phastCons77way and update for this # situation ln -s `pwd`/* \ /usr/local/apache/htdocs-hgdownload/goldenPath/regenRn1/phastCons7way ##################################################################### cd /hive/data/genomes/regenRn1/bed/multiz7way/downloads/phyloP7way ln -s ../../consPhyloP/all/downloads/phyloP7way.wigFix.gz \ ./regenRn1.phyloP7way.wigFix.gz ln -s ../../consPhyloP/run.phyloP/all.mod regenRn1.phyloP7way.mod ln -s ../../consPhyloP/all/phyloP7way.bw regenRn1.phyloP7way.bw time md5sum *.mod *.bw *.gz > md5sum.txt # real 0m18.237s # obtain the README.txt from galGal6/phyloP77way and update for this # situation ln -s `pwd`/* \ /usr/local/apache/htdocs-hgdownload/goldenPath/regenRn1/phyloP7way ########################################################################### ## create upstream refGene maf files cd /hive/data/genomes/regenRn1/bed/multiz7way/downloads/multiz7way # bash script #!/bin/sh export geneTbl="refGene" for S in 1000 2000 5000 do echo "making upstream${S}.maf" featureBits regenRn1 ${geneTbl}:upstream:${S} -fa=/dev/null -bed=stdout \ | perl -wpe 's/_up[^\t]+/\t0/' | sort -k1,1 -k2,2n \ | /cluster/bin/$MACHTYPE/mafFrags regenRn1 multiz7way \ stdin stdout \ -orgs=/hive/data/genomes/regenRn1/bed/multiz7way/species.list \ | gzip -c > upstream${S}.${geneTbl}.maf.gz echo "done upstream${S}.${geneTbl}.maf.gz" done XXX - running - Thu Oct 17 15:09:16 PDT 2019 # about 10 minutes md5sum *.maf.gz *.nh upstream*.gz README.txt >> md5sum.txt # some other symlinks were already made above # obtain the README.txt from cavPor3/multiz7way and update for this # situation ln -s `pwd`/upstream*.gz `pwd`/README.txt \ /usr/local/apache/htdocs-hgdownload/goldenPath/regenRn1/multiz7way ############################################################################# # hgPal downloads (TBD - 2016-06-11 - Hiram) # FASTA from 7way for knownGene, refGene and knownCanonical ssh hgwdev screen -S regenRn1HgPal mkdir /hive/data/genomes/regenRn1/bed/multiz7way/pal cd /hive/data/genomes/regenRn1/bed/multiz7way/pal cat ../species.list | tr '[ ]' '[\n]' > order.list # this for loop takes about 2.6 hours on this large count contig assembly export mz=multiz7way export gp=refGene export db=regenRn1 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 176m60.376s export mz=multiz7way export gp=refGene time find ./exonAA -type f | grep exonAA.fa.gz | xargs zcat \ | gzip -c > $gp.$mz.exonAA.fa.gz # real 10m29.600s time find ./exonNuc -type f | grep exonNuc.fa.gz | xargs zcat \ | gzip -c > $gp.$mz.exonNuc.fa.gz # real 16m9.974s # -rw-rw-r-- 1 611281644 Apr 16 20:37 refGene.multiz7way.exonAA.fa.gz # -rw-rw-r-- 1 966671426 Apr 16 21:06 refGene.multiz7way.exonNuc.fa.gz export mz=multiz7way export gp=refGene export db=regenRn1 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 7way (DONE - 2017-12-18 - Hiram) mkdir /hive/users/hiram/bigWays/regenRn1.7way cd /hive/users/hiram/bigWays echo "regenRn1" > regenRn1.7way/ordered.list awk '{print $1}' /hive/data/genomes/regenRn1/bed/multiz7way/7way.distances.txt \ >> regenRn1.7way/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 regenRn1.7way/ordered.list # dbDb.sh constructs regenRn1.7way/GalVar1_7way_conservation_alignment.html # may need to add new assembly references to srcReference.list and # urlReference.list ./dbDb.sh regenRn1 7way # sizeStats.pl constructs regenRn1.7way/GalVar1_7way_Genome_size_statistics.html # this requires entries in coverage.list for new sequences ./sizeStats.pl regenRn1 7way # defCheck.pl constructs GalVar1_7way_conservation_lastz_parameters.html ./defCheck.pl regenRn1 7way # this constructs the html pages in regenRn1.7way/: # -rw-rw-r-- 3848 Dec 18 14:05 GalVar1_7way_conservation_alignment.html # -rw-rw-r-- 5500 Dec 18 14:05 GalVar1_7way_Genome_size_statistics.html # -rw-rw-r-- 3613 Dec 18 14:05 GalVar1_7way_conservation_lastz_parameters.html # add those pages to the genomewiki. Their page names are the # names of the .html files without the .html: # GalVar1_7way_conservation_alignment # GalVar1_6way_Genome_size_statistics # GalVar1_6way_conservation_lastz_parameters # when you view the first one you enter, it will have links to the # missing two. ############################################################################