From afbf9340a13f6340f7cb104ae29e5b50b19162f6 Mon Sep 17 00:00:00 2001 From: David Jones Date: Thu, 24 Nov 2016 16:13:17 +0000 Subject: [PATCH 01/26] Added diff_bams as a c program. 2hrs(perl)->45mins(c) for a 68G bam file --- c/Makefile | 10 +- c/diff_bams.c | 302 ++++++++++++++++++++++++++++++++++++++++++++++++++ setup.sh | 1 + 3 files changed, 310 insertions(+), 3 deletions(-) create mode 100644 c/diff_bams.c diff --git a/c/Makefile b/c/Makefile index ce223f4..ae1979b 100644 --- a/c/Makefile +++ b/c/Makefile @@ -55,6 +55,7 @@ MD := mkdir #Build target executable BAM_STATS_TARGET=../bin/bam_stats SQ_TARGET=../bin/reheadSQ +BAM_DIFF=../bin/diff_bams # # The following part of the makefile is generic; it can be used to @@ -66,7 +67,7 @@ SQ_TARGET=../bin/reheadSQ .NOTPARALLEL: test -all: clean pre make_htslib_tmp $(BAM_STATS_TARGET) $(BAM2BG_TARGET) $(BAM2BW_TARGET) test remove_htslib_tmp $(CAT_TARGET) $(SQ_TARGET) +all: clean pre make_htslib_tmp $(BAM_STATS_TARGET) $(BAM2BG_TARGET) $(BAM2BW_TARGET) $(BAM_DIFF) test remove_htslib_tmp $(CAT_TARGET) $(SQ_TARGET) @echo bam_stats and reheadSQ compiled. $(BAM_STATS_TARGET): $(OBJS) @@ -75,6 +76,9 @@ $(BAM_STATS_TARGET): $(OBJS) $(SQ_TARGET): $(CC) $(CFLAGS) ./reheadSQ.c -o $(SQ_TARGET) +$(BAM_DIFF): + $(CC) $(CFLAGS) $(INCLUDES) -o $(BAM_DIFF) $(OBJS) $(LFLAGS) $(LIBS) ./diff_bams.c + #Unit Tests test: $(BAM_STATS_TARGET) @@ -97,7 +101,7 @@ remove_htslib_tmp: copyscript: cp ./scripts/* ./bin/ - chmod a+x $(BAM_STATS_TARGET) $(CAT_TARGET) $(SQ_TARGET) $(BAM2BW_TARGET) $(BAM2BG_TARGET) + chmod a+x $(BAM_STATS_TARGET) $(CAT_TARGET) $(SQ_TARGET) $(BAM2BW_TARGET) $(BAM2BG_TARGET) $(BAM_DIFF) valgrind: VALGRIND="valgrind --log-file=/tmp/valgrind-%p.log" $(MAKE) @@ -112,7 +116,7 @@ valgrind: clean: @echo clean - $(RM) ./*.o *~ $(BAM_STATS_TARGET) $(SQ_TARGET) ./tests/tests_log $(TESTS) ./*.gcda ./*.gcov ./*.gcno *.gcda *.gcov *.gcno ./tests/*.gcda ./tests/*.gcov ./tests/*.gcno + $(RM) ./*.o *~ $(BAM_STATS_TARGET) $(SQ_TARGET) $(BAM_DIFF) ./tests/tests_log $(TESTS) ./*.gcda ./*.gcov ./*.gcno *.gcda *.gcov *.gcno ./tests/*.gcda ./tests/*.gcov ./tests/*.gcno -rm -rf $(HTSTMP) depend: $(SRCS) diff --git a/c/diff_bams.c b/c/diff_bams.c new file mode 100644 index 0000000..0a943c3 --- /dev/null +++ b/c/diff_bams.c @@ -0,0 +1,302 @@ +#include +#include +#include "htslib/sam.h" +#include "khash.h" +#include "dbg.h" + +KHASH_MAP_INIT_INT(posn,int32_t) +KHASH_MAP_INIT_INT(chrom,khash_t(posn)) + +char *bam_a_loc = NULL; +char *bam_b_loc = NULL; +char *ref_file = NULL; +int skip_z = 0; +int count_flag_diff = 0; + + +int check_exist(char *fname){ + FILE *fp; + if((fp = fopen(fname,"r"))){ + fclose(fp); + return 1; + } + return 0; +} + +void print_version (int exit_code){ + printf ("%s\n",VERSION); + exit(exit_code); +} + +void print_usage (int exit_code){ + + printf ("Usage: diff_bams -a bam_a.bam -b bam_b.bsm [-r reference.fa] [] [] [-h] [-v]\n\n"); + printf ("Required:\n"); + printf ("-a --bam_a The first BAM|CRAM file.\n"); + printf ("-b --bam_b The second BAM|CRAM file.\n\n"); + printf ("Other:\n"); + printf ("-r --ref Required for CRAM, genome.fa with co-located fai.\n"); + printf ("-c --count Count flag differences.\n"); + printf ("-s --skip Don't include reads with MAPQ=0 in comparison.\n\n"); + printf ("-h --help Display this usage information.\n"); + printf ("-v --version Prints the version number.\n\n"); + exit(exit_code); +} + +void options(int argc, char *argv[]){ + + const struct option long_opts[] = + { + {"version", no_argument, 0, 'v'}, + {"help",no_argument,0,'h'}, + {"bam_a",required_argument,0,'a'}, + {"ref",required_argument,0,'r'}, + {"bam_b",required_argument,0,'b'}, + {"skip",no_argument,0,'s'}, + {"count",no_argument,0,'c'}, + { NULL, 0, NULL, 0} + + }; //End of declaring opts + + int index = 0; + int iarg = 0; + + //Iterate through options + while((iarg = getopt_long(argc, argv, "a:b:r:scvh", long_opts, &index)) != -1){ + switch(iarg){ + case 's': + skip_z = 1; + break; + + case 'c': + count_flag_diff = 1; + break; + + case 'r': + ref_file = optarg; + break; + + case 'a': + bam_a_loc = optarg; + break; + + case 'b': + bam_b_loc = optarg; + break; + + case 'h': + print_usage(0); + break; + + case 'v': + print_version(0); + break; + + case '?': + print_usage (1); + break; + + default: + print_usage (1); + + }; // End of args switch statement + + }//End of iteration through options + + //Do some checking to ensure required arguments were passed and are accessible files + if(ref_file != NULL){ + if(check_exist(ref_file) != 1){ + fprintf(stderr,"Reference fasta file (-r) %s does not exist.\n",ref_file); + print_usage(1); + } + } + + if(strcmp(bam_a_loc,bam_b_loc)==0){ + fprintf(stderr,"bam_a '%s' and bam_b '%s' cannot be the same file\n",bam_a_loc,bam_b_loc; + print_usage(1); + } + + if(check_exist(bam_a_loc) != 1){ + fprintf(stderr,"First BAM|CRAM file (-a) %s does not exist.\n",bam_a_loc); + print_usage(1); + } + + if(check_exist(bam_b_loc) != 1){ + fprintf(stderr,"Second BAM|CRAM file (-b) %s does not exist.\n",bam_b_loc); + print_usage(1); + } + + if((strcmp(bam_a_loc + strlen(bam_a_loc) - 4,".cram")==0 || strcmp(bam_b_loc + strlen(bam_b_loc) - 4,".cram")==0) && ref_file == NULL) { + fprintf(stderr,"Option '-r' must be defined if any CRAM files are provided\n"); + print_usage(1); + } + + return; +} + +int main(int argc, char *argv[]){ + htsFile *htsa = NULL; + htsFile *htsb = NULL; + bam_hdr_t *heada = NULL; + bam_hdr_t *headb = NULL; + khash_t(chrom) *chr_hash = NULL; + bam1_t *reada = NULL; + bam1_t *readb = NULL; + options(argc, argv); + //Open bam file a + htsa = hts_open(bam_a_loc,"r"); + check(htsa != NULL, "Error opening hts file 'a' for reading '%s'.",bam_a_loc); + //Open bam file b + htsb = hts_open(bam_b_loc,"r"); + check(htsb != NULL, "Error opening hts file 'b' for reading '%s'.",bam_b_loc); + + if(ref_file){ + hts_set_fai_filename(htsa, ref_file); + hts_set_fai_filename(htsb, ref_file); + } + + heada = sam_hdr_read(htsa); + check(heada != NULL, "Error reading header from opened hts file 'a' '%s'.",bam_a_loc); + headb = sam_hdr_read(htsb); + check(headb != NULL, "Error reading header from opened hts file 'b' '%s'.",bam_b_loc); + + if(heada->n_targets != headb->n_targets ){ + sentinel("Reference sequence count is different\n"); + } + fprintf(stdout,"Reference sequence count passed\n"); + + int i=0; + for(i=0;in_targets;i++){ + if(strcmp(heada->target_name[i],headb->target_name[i])!=0){ + sentinel("Reference sequences in different order\n"); + } + } + fprintf(stdout,"Reference sequence order passed\n"); + + uint64_t count = 0; + uint64_t flag_diffs = 0; + uint64_t last_coord = 0; + int chka = 0; + int chkb = 0; + reada = bam_init1(); + readb = bam_init1(); + while(1){ + count++; + //Check the individual reads + chka = sam_read1(htsa,heada,reada); + chkb = sam_read1(htsb,headb,readb); + + if(skip_z==1){ + //Move to next non 0 MQ read + while(reada->core.qual == 0 && chka >= 0) { + chka = sam_read1(htsa,heada,reada); + } + while(readb->core.qual == 0 && chkb >= 0) { + chkb = sam_read1(htsb,headb,readb); + } + } + + if(chka<0 && chkb<0){ + break; + } + if((chka>=0 && chkb <0) || (chkb>=0 && chka<0)){ + sentinel("Files have different number of records\n"); + } + + if(reada->core.tid != readb->core.tid || reada->core.pos != readb->core.pos || strcmp(bam_get_qname(reada),bam_get_qname(readb))!=0){ + sentinel("Files differ at record %"PRIu64" (qname) a=%s b=%s\n",count,bam_get_qname(reada),bam_get_qname(readb)); + } + + if(reada->core.flag != readb->core.flag){ + if(count_flag_diff==1){ + if(chr_hash == NULL){ chr_hash = kh_init(chrom);} + flag_diffs++; + int32_t pos = reada->core.pos; + if((pos - last_coord) >=0 ){ //Checking for different chr + int res; + khiter_t k = kh_get(chrom, chr_hash, reada->core.tid); // query the hash table + khash_t(posn) *pos_hash = NULL; + if(k == kh_end(chr_hash)){ + k = kh_put(chrom,chr_hash,reada->core.tid,&res); + pos_hash = kh_init(posn); + khiter_t kpos = kh_put(posn,pos_hash,pos,&res); + kh_value(pos_hash,kpos) = 1; + kh_value(chr_hash,k) = *pos_hash; + }else{ + *pos_hash = kh_value(chr_hash,k); + khiter_t kpos = kh_get(posn, pos_hash, pos); // query the pos hash table + if(kpos == kh_end(pos_hash)){ + kpos = kh_put(posn,pos_hash,pos,&res); + kh_value(pos_hash,kpos) = 1; + }else{ + kh_value(pos_hash,kpos) = kh_value(pos_hash,kpos)+1; + } + kh_value(chr_hash,k) = *pos_hash; + } + } + last_coord = pos; + }else{ + sentinel("Files differ at record %"PRIu64" (flags) a=%s b=%s\n",count,bam_get_qname(reada),bam_get_qname(readb)); + } + }//End of if flags don't match + if(count % 5000000 == 0) { + fprintf(stdout,"Matching records: %"PRIu64"",count); + if(count_flag_diff){ + fprintf(stdout,"\t(flag mismatch: %"PRIu64")",flag_diffs); + } + fprintf(stdout,"\r"); + } + }//End of looping through all reads + + fprintf(stdout,"Matching records: %"PRIu64"\n",count); + if(count_flag_diff && chr_hash != NULL){ + fprintf(stdout,"Flag mismatches: %"PRIu64"\n",flag_diffs); + fprintf(stdout,"Locations of flag differences:\n"); + fprintf(stdout,"#Chr\tPos\tCount\n"); + khiter_t k; + for (k = kh_begin(chr_hash); k != kh_end(chr_hash); ++k) { + if (kh_exist(chr_hash, k)) { + const int32_t keyp = kh_key(chr_hash,k); + khash_t(posn) pos_hash = kh_value(chr_hash, k); + khiter_t kpos; + for (kpos = kh_begin(&pos_hash); kpos != kh_end(&pos_hash); ++kpos) { + if (kh_exist(&pos_hash, kpos)) { + const int32_t key = kh_key(&pos_hash,kpos); + int32_t counter = kh_value(&pos_hash,kpos); + fprintf(stdout,"%s\t%"PRIi32"\t%"PRIi32"\n",heada->target_name[keyp], key, counter); + kh_destroy(posn,&pos_hash); + } + } + + } + } + kh_destroy(chrom,chr_hash); + } + + bam_destroy1(reada); + bam_destroy1(readb); + bam_hdr_destroy(heada); + bam_hdr_destroy(headb); + hts_close(htsa); + hts_close(htsb); + return 0; +error: + if(count_flag_diff && chr_hash != NULL){ + khiter_t k; + for (k = kh_begin(chr_hash); k != kh_end(chr_hash); ++k) { + if (kh_exist(chr_hash, k)) { + const int32_t keyp = kh_key(chr_hash,k); + khash_t(posn) pos_hash = kh_value(chr_hash, k); + kh_destroy(posn,&pos_hash); + } + } + kh_destroy(chrom,chr_hash); + } + if(reada) bam_destroy1(reada); + if(readb) bam_destroy1(readb); + if(heada) bam_hdr_destroy(heada); + if(headb) bam_hdr_destroy(headb); + if(htsa) hts_close(htsa); + if(htsb) hts_close(htsb); + return 1; +} \ No newline at end of file diff --git a/setup.sh b/setup.sh index 49a0be5..7801985 100755 --- a/setup.sh +++ b/setup.sh @@ -285,6 +285,7 @@ else env HTSLIB=$SETUP_DIR/htslib make -C c -j$CPU prefix=$INST_PATH cp bin/bam_stats $INST_PATH/bin/. cp bin/reheadSQ $INST_PATH/bin/. + cp bin/diff_bams $INST_PATH/bin/. touch $SETUP_DIR/bam_stats.success make -C c clean fi From 4c688bd44389f576ee1dbf389594bab5eede2fdb Mon Sep 17 00:00:00 2001 From: David Jones Date: Thu, 24 Nov 2016 16:26:40 +0000 Subject: [PATCH 02/26] Corrected missing ( --- c/diff_bams.c | 2 +- 1 file changed, 1 insertion(+), 1 deletion(-) diff --git a/c/diff_bams.c b/c/diff_bams.c index 0a943c3..2474791 100644 --- a/c/diff_bams.c +++ b/c/diff_bams.c @@ -112,7 +112,7 @@ void options(int argc, char *argv[]){ } if(strcmp(bam_a_loc,bam_b_loc)==0){ - fprintf(stderr,"bam_a '%s' and bam_b '%s' cannot be the same file\n",bam_a_loc,bam_b_loc; + fprintf(stderr,"bam_a '%s' and bam_b '%s' cannot be the same file\n",bam_a_loc,bam_b_loc); print_usage(1); } From b7feabc5b4ead19f81a30deeb79fa8952b5efb6d Mon Sep 17 00:00:00 2001 From: Keiran Raine Date: Fri, 13 Jan 2017 15:01:58 +0000 Subject: [PATCH 03/26] Add initial RNA download code --- bin/gnos_pull.pl | 58 ++++++++++++++++++++++++++++++++++++++++-------- 1 file changed, 49 insertions(+), 9 deletions(-) diff --git a/bin/gnos_pull.pl b/bin/gnos_pull.pl index eab9cf6..b536b1c 100755 --- a/bin/gnos_pull.pl +++ b/bin/gnos_pull.pl @@ -2,7 +2,7 @@ ##########LICENCE########## # PCAP - NGS reference implementations and helper code for the ICGC/TCGA Pan-Cancer Analysis Project -# Copyright (C) 2014-2015 ICGC PanCancer Project +# Copyright (C) 2014-2017 ICGC PanCancer Project # # This program is free software; you can redistribute it and/or # modify it under the terms of the GNU General Public License @@ -49,7 +49,7 @@ use PCAP; use PCAP::Cli; -const my @ANALYSIS_TYPES => (qw(ALIGNMENTS CALLS)); +const my @ANALYSIS_TYPES => (qw(ALIGNMENTS CALLS RNA_STAR RNA_TOPHAT)); const my @AVAILABLE_COMPOSITE_FILTERS => (qw(not_sanger_workflow caller max_dataset_GB multi_tumour sanger_version broad_version dkfz_embl_version jamboree_approved manual_donor_blacklist)); const my $DEFAULT_URL => 'http://pancancer.info/gnos_metadata/latest'; const my $GTDL_COMMAND => '%s%s --max-children 3 --rate-limit 200 -vv -c %s -d %scghub/data/analysis/download/%s -p %s'; @@ -152,6 +152,10 @@ sub pull_data { $check_ref = \&check_alignments; $code_ref = \&pull_alignments; } + elsif($options->{'analysis'} =~ m/^RNA/) { + $check_ref = \&check_rna_alignments; + $code_ref = \&pull_rna_alignments; + } my $thread_count = $options->{'threads'}; if($CAN_USE_THREADS == 0) { @@ -218,6 +222,35 @@ sub check_or_create_symlink { return 1; } +sub check_rna_alignments{ + my ($options, $donor, $outbase, $donor_base) = @_; + warn "Checking $donor->{donor_unique_id}\n"; + $options->{'analysis'} =~ m/^RNA_(.+)/; + my $rna_type = lc $1; + my $to_do = 0; + # for normal: + $to_do += check_bam($options, $donor->{'donor_unique_id'}, $donor->{'rna_seq'}->{'alignment'}->{'normal'}->{$rna_type}, $outbase, $donor_base, 'rna/'.$rna_type.'/normal'); + + # for tumour + for my $tumour_data(@{$donor->{'rna_seq'}->{'alignment'}->{'tumor'}}) { + $to_do += check_bam($options, $donor->{'donor_unique_id'}, $tumour_data->{$rna_type}, $outbase, $donor_base, 'rna/'.$rna_type.'/tumour'); + } + return $to_do; +} + +sub pull_rna_bam { + my ($options, $donor, $outbase, $donor_base) = @_; + $options->{'analysis'} =~ m/^RNA_(.+)/; + my $rna_type = lc $1; + # for normal: + pull_bam($options, $donor->{'donor_unique_id'}, $donor->{'rna_seq'}->{'alignment'}->{'normal'}->{$rna_type}, $outbase, $donor_base, 'rna/'.$rna_type.'/normal'); + + # for tumour + for my $tumour_data(@{$donor->{'tumor_alignment_status'}}) { + pull_bam($options, $donor->{'donor_unique_id'}, $tumour_data->{$rna_type}, $outbase, $donor_base, 'rna/'.$rna_type.'/tumour'); + } +} + sub check_alignments { my ($options, $donor, $outbase, $donor_base) = @_; warn "Checking $donor->{donor_unique_id}\n"; @@ -516,13 +549,15 @@ sub load_data { warn sprintf "Donor: %s blacklisted\n", $donor->{'donor_unique_id'} if($options->{'debug'}); next; } - unless($donor->{'flags'}->{'is_normal_specimen_aligned'}) { - warn sprintf "Donor: %s normal sample not aligned\n", $donor->{'donor_unique_id'} if($options->{'debug'}); - next; - } - unless($donor->{'flags'}->{'are_all_tumor_specimens_aligned'}) { - warn sprintf "Donor: %s all tumour samples not aligned\n", $donor->{'donor_unique_id'} if($options->{'debug'}); - next; + unless($options->{'analysis'} =~ m/^RNA/) { + unless($donor->{'flags'}->{'is_normal_specimen_aligned'}) { + warn sprintf "Donor: %s normal sample not aligned\n", $donor->{'donor_unique_id'} if($options->{'debug'}); + next; + } + unless($donor->{'flags'}->{'are_all_tumor_specimens_aligned'}) { + warn sprintf "Donor: %s all tumour samples not aligned\n", $donor->{'donor_unique_id'} if($options->{'debug'}); + next; + } } if(exists $options->{'COMPOSITE_FILTERS'}->{'multi_tumour'} && $donor->{'flags'}->{'all_tumor_specimen_aliquot_counts'} == 1) { @@ -574,6 +609,11 @@ sub load_data { } } + elsif($options->{'analysis'} =~ m/^RNA_(.+)/) { + my $rna_type = lc $1; + next unless( $donor->{'flags'}->{'is_tumor_'.$rna_type.'_rna_seq_alignment_performed'} + || $donor->{'flags'}->{'is_normal_'.$rna_type.'_rna_seq_alignment_performed'}); + } else { my $size = data_size_alignments_gb($options, $donor); if(exists $options->{'COMPOSITE_FILTERS'}->{'max_dataset_GB'} && $size > $options->{'COMPOSITE_FILTERS'}->{'max_dataset_GB'}) { From efbd34852655bbdf81a1f24c18e3591ca9d23e13 Mon Sep 17 00:00:00 2001 From: Keiran Raine Date: Fri, 13 Jan 2017 15:56:06 +0000 Subject: [PATCH 04/26] Correct function name --- bin/gnos_pull.pl | 2 +- 1 file changed, 1 insertion(+), 1 deletion(-) diff --git a/bin/gnos_pull.pl b/bin/gnos_pull.pl index b536b1c..067c8cd 100755 --- a/bin/gnos_pull.pl +++ b/bin/gnos_pull.pl @@ -238,7 +238,7 @@ sub check_rna_alignments{ return $to_do; } -sub pull_rna_bam { +sub pull_rna_alignments { my ($options, $donor, $outbase, $donor_base) = @_; $options->{'analysis'} =~ m/^RNA_(.+)/; my $rna_type = lc $1; From f60d96662b5cb6d4b74c696e440ba92bdb60c21c Mon Sep 17 00:00:00 2001 From: Keiran Raine Date: Wed, 25 Jan 2017 16:30:34 +0000 Subject: [PATCH 05/26] Adds RNA download support --- bin/gnos_pull.pl | 25 ++++++++++++++----------- 1 file changed, 14 insertions(+), 11 deletions(-) diff --git a/bin/gnos_pull.pl b/bin/gnos_pull.pl index 067c8cd..2a92ff5 100755 --- a/bin/gnos_pull.pl +++ b/bin/gnos_pull.pl @@ -229,7 +229,9 @@ sub check_rna_alignments{ my $rna_type = lc $1; my $to_do = 0; # for normal: - $to_do += check_bam($options, $donor->{'donor_unique_id'}, $donor->{'rna_seq'}->{'alignment'}->{'normal'}->{$rna_type}, $outbase, $donor_base, 'rna/'.$rna_type.'/normal'); + if(exists $donor->{'rna_seq'}->{'alignment'}->{'normal'}->{$rna_type}) { + $to_do += check_bam($options, $donor->{'donor_unique_id'}, $donor->{'rna_seq'}->{'alignment'}->{'normal'}->{$rna_type}, $outbase, $donor_base, 'rna/'.$rna_type.'/normal'); + } # for tumour for my $tumour_data(@{$donor->{'rna_seq'}->{'alignment'}->{'tumor'}}) { @@ -243,10 +245,12 @@ sub pull_rna_alignments { $options->{'analysis'} =~ m/^RNA_(.+)/; my $rna_type = lc $1; # for normal: - pull_bam($options, $donor->{'donor_unique_id'}, $donor->{'rna_seq'}->{'alignment'}->{'normal'}->{$rna_type}, $outbase, $donor_base, 'rna/'.$rna_type.'/normal'); + if(exists $donor->{'rna_seq'}->{'alignment'}->{'normal'}->{$rna_type}) { + pull_bam($options, $donor->{'donor_unique_id'}, $donor->{'rna_seq'}->{'alignment'}->{'normal'}->{$rna_type}, $outbase, $donor_base, 'rna/'.$rna_type.'/normal'); + } # for tumour - for my $tumour_data(@{$donor->{'tumor_alignment_status'}}) { + for my $tumour_data(@{$donor->{'rna_seq'}->{'alignment'}->{'tumor'}}) { pull_bam($options, $donor->{'donor_unique_id'}, $tumour_data->{$rna_type}, $outbase, $donor_base, 'rna/'.$rna_type.'/tumour'); } } @@ -287,8 +291,8 @@ sub check_bam { if(-e $success) { check_or_create_symlink($orig_bam, $sym_bam); - check_or_create_symlink($orig_bam.'.bai', $sym_bam.'.bai'); - create_bas($repo, $gnos_id, $sym_bam); + check_or_create_symlink($orig_bam.'.bai', $sym_bam.'.bai') if(-e $orig_bam.'.bai'); + create_bas($repo, $gnos_id, $sym_bam) unless($options->{'analysis'} =~ m/^RNA/); return 0; } @@ -310,7 +314,6 @@ sub pull_alignments { sub pull_bam { my ($options, $donor_id, $bam_data, $outbase, $donor_base, $type) = @_; - my $repo = select_repo($options, $bam_data->{'aligned_bam'}->{'gnos_repo'}); unless(exists $options->{'keys'}->{$repo}) { warn sprintf "Skipping %s BAM for Donor %s - No permission key for repo %s", $type, $donor_id, $repo; @@ -330,8 +333,8 @@ sub pull_bam { if(-e $success) { check_or_create_symlink($orig_bam, $sym_bam); - check_or_create_symlink($orig_bam.'.bai', $sym_bam.'.bai'); - create_bas($repo, $gnos_id, $sym_bam); + check_or_create_symlink($orig_bam.'.bai', $sym_bam.'.bai') if(-e $orig_bam.'.bai'); + create_bas($repo, $gnos_id, $sym_bam) unless($options->{'analysis'} =~ m/^RNA/); return; } return if($options->{'symlinks'}); @@ -362,8 +365,9 @@ sub pull_bam { unlink $err_file; check_or_create_symlink($orig_bam, $sym_bam); - check_or_create_symlink($orig_bam.'.bai', $sym_bam.'.bai'); - my $bas_valid = create_bas($repo, $gnos_id, $sym_bam); + check_or_create_symlink($orig_bam.'.bai', $sym_bam.'.bai') if(-e $orig_bam.'.bai'); + my $bas_valid = 1; + $bas_valid = create_bas($repo, $gnos_id, $sym_bam) unless($options->{'analysis'} =~ m/^RNA/); if($bas_valid == 1) { # touch a success file in the output loc @@ -607,7 +611,6 @@ sub load_data { warn sprintf "Donor: %s has no variant calling available\n", $donor->{'donor_unique_id'} if($options->{'debug'}); next; } - } elsif($options->{'analysis'} =~ m/^RNA_(.+)/) { my $rna_type = lc $1; From 17846c19b070a9826f50fb6f6c332ec8977132ba Mon Sep 17 00:00:00 2001 From: Keiran Raine Date: Wed, 25 Jan 2017 18:22:57 +0000 Subject: [PATCH 06/26] Change to running script as bash input instead of chmod to save file operations --- lib/PCAP/Threaded.pm | 6 ++---- 1 file changed, 2 insertions(+), 4 deletions(-) diff --git a/lib/PCAP/Threaded.pm b/lib/PCAP/Threaded.pm index 6d0470a..6762817 100644 --- a/lib/PCAP/Threaded.pm +++ b/lib/PCAP/Threaded.pm @@ -2,7 +2,7 @@ package PCAP::Threaded; ##########LICENCE########## # PCAP - NGS reference implementations and helper code for the ICGC/TCGA Pan-Cancer Analysis Project -# Copyright (C) 2014 ICGC PanCancer Project +# Copyright (C) 2014-2017 ICGC PanCancer Project # # This program is free software; you can redistribute it and/or # modify it under the terms of the GNU General Public License @@ -224,7 +224,7 @@ sub external_process_handler { my $err = File::Spec->catfile($tmp, "$caller.$suffix.err"); try { - system("/usr/bin/time $script 1> $out 2> $err"); + system("/usr/bin/time bash $script 1> $out 2> $err"); } catch { die $_; }; @@ -244,8 +244,6 @@ sub _create_script { print $SH qq{\n}; close $SH; sleep 1; - chmod $SCRIPT_OCT_MODE, $script; - sleep 1; return $script; } From d31190b48e103888cea94b87a610e9a5436bee1d Mon Sep 17 00:00:00 2001 From: Keiran Raine Date: Wed, 25 Jan 2017 19:31:56 +0000 Subject: [PATCH 07/26] Addition of function to confirm that files have completed write when to be used in execution following some issues on high CPU hosts. Testing of the function was manual as a test case would require more code than the function. If modifying this will show it functions: Without creating the file /tmp/thisIsFun execute (debug indicated): ``` rm -f /tmp/thisIsFun perl -I lib -e 'use PCAP::Threaded; PCAP::Threaded::_file_complete("/tmp/thisIsFun", 1);' ``` This should exit in ~15 seconds showing that the file wasn't present. ``` touch /tmp/thisIsFun perl -I lib -e 'use PCAP::Threaded; PCAP::Threaded::_file_complete("/tmp/thisIsFun", 1);' ``` Will exit instantly as file is found. Now test the blocking if the file is continuously written to: ``` /usr/bin/yes > /tmp/thisIsFun perl -I lib -e 'use PCAP::Threaded; PCAP::Threaded::_file_complete("/tmp/thisIsFun", 1);' ``` This will exit in < 15 seconds, but due to the file being written to rather than absence. Finally repeat the above, but killing the `yes` command after starting the perl will cause the perl process to complete as the file is closed. --- lib/PCAP/Threaded.pm | 29 ++++++++++++++++++++++++++--- 1 file changed, 26 insertions(+), 3 deletions(-) diff --git a/lib/PCAP/Threaded.pm b/lib/PCAP/Threaded.pm index 6762817..8ac6dae 100644 --- a/lib/PCAP/Threaded.pm +++ b/lib/PCAP/Threaded.pm @@ -34,12 +34,11 @@ use Capture::Tiny qw(capture); use IO::File; use Const::Fast qw(const); use Scalar::Util qw(looks_like_number); +use Time::HiRes qw(usleep); our $CAN_USE_THREADS = 0; $CAN_USE_THREADS = eval 'use threads; 1'; -const my $SCRIPT_OCT_MODE => 0777; - our $OUT_ERR = 1; sub new { @@ -243,10 +242,34 @@ sub _create_script { print $SH join qq{\n}, @{$commands}; print $SH qq{\n}; close $SH; - sleep 1; + _file_complete($script); return $script; } +sub _file_complete { + my ($script, $debug) = @_; + my $tries = 0; + my $millisec; + while(1) { + croak "Failed to confirm script written after 30 attempts: $script" if($tries > 30); + my ($stdout, $stderr, $exit) = capture { system([0,1], 'lsof', $script); }; + if($exit > 1) { + croak $stderr; + } + printf STDERR "OUT : %s\nERR : %s\nEXIT: %s\n", $stdout,$stderr,$exit if($debug); + if($exit == 1) { + last if(-e $script); + } + # sleep for random millisec max 0.5 sec. + $millisec = rand 500_000; + $millisec = 10000 if($millisec < 10000); # need to allow something to happen + warn $millisec if($debug); + usleep($millisec); + $tries++; + } + return 1; +} + 1; __END__ From 91454b13c4e8f1c94b89cd7ac8e53049a105021b Mon Sep 17 00:00:00 2001 From: Keiran Raine Date: Thu, 26 Jan 2017 06:23:12 +0000 Subject: [PATCH 08/26] switch back to executable file --- lib/PCAP/Threaded.pm | 19 ++++++++++++------- 1 file changed, 12 insertions(+), 7 deletions(-) diff --git a/lib/PCAP/Threaded.pm b/lib/PCAP/Threaded.pm index 8ac6dae..50a464e 100644 --- a/lib/PCAP/Threaded.pm +++ b/lib/PCAP/Threaded.pm @@ -39,6 +39,8 @@ use Time::HiRes qw(usleep); our $CAN_USE_THREADS = 0; $CAN_USE_THREADS = eval 'use threads; 1'; +const my $SCRIPT_OCT_MODE => 0777; + our $OUT_ERR = 1; sub new { @@ -223,7 +225,7 @@ sub external_process_handler { my $err = File::Spec->catfile($tmp, "$caller.$suffix.err"); try { - system("/usr/bin/time bash $script 1> $out 2> $err"); + system("/usr/bin/time $script 1> $out 2> $err"); } catch { die $_; }; @@ -242,14 +244,20 @@ sub _create_script { print $SH join qq{\n}, @{$commands}; print $SH qq{\n}; close $SH; - _file_complete($script); + + # sleep for random millisec max 1 sec. + my $millisec = rand 1_000_000; + $millisec = 100_000 if($millisec < 100_000); # min 0.1 sec + + _file_complete($script, $millisec); + chmod $SCRIPT_OCT_MODE, $script; + usleep($millisec); return $script; } sub _file_complete { - my ($script, $debug) = @_; + my ($script, $millisec, $debug) = @_; my $tries = 0; - my $millisec; while(1) { croak "Failed to confirm script written after 30 attempts: $script" if($tries > 30); my ($stdout, $stderr, $exit) = capture { system([0,1], 'lsof', $script); }; @@ -260,9 +268,6 @@ sub _file_complete { if($exit == 1) { last if(-e $script); } - # sleep for random millisec max 0.5 sec. - $millisec = rand 500_000; - $millisec = 10000 if($millisec < 10000); # need to allow something to happen warn $millisec if($debug); usleep($millisec); $tries++; From 2ca68c10cc9f6cba32cd00a86bccd0d9c1dbac6a Mon Sep 17 00:00:00 2001 From: Keiran Raine Date: Thu, 26 Jan 2017 06:24:20 +0000 Subject: [PATCH 09/26] Everything possible to ensure fh not active --- lib/PCAP/Threaded.pm | 1 + 1 file changed, 1 insertion(+) diff --git a/lib/PCAP/Threaded.pm b/lib/PCAP/Threaded.pm index 50a464e..66081d7 100644 --- a/lib/PCAP/Threaded.pm +++ b/lib/PCAP/Threaded.pm @@ -244,6 +244,7 @@ sub _create_script { print $SH join qq{\n}, @{$commands}; print $SH qq{\n}; close $SH; + undef $SH; # sleep for random millisec max 1 sec. my $millisec = rand 1_000_000; From bf00781454c359741a13e728a0c086123ab8c8f6 Mon Sep 17 00:00:00 2001 From: Keiran Raine Date: Fri, 27 Jan 2017 10:42:27 +0000 Subject: [PATCH 10/26] hardened script exec, debug forced for now --- lib/PCAP/Threaded.pm | 40 ++++++++++++++++++++++------------------ 1 file changed, 22 insertions(+), 18 deletions(-) diff --git a/lib/PCAP/Threaded.pm b/lib/PCAP/Threaded.pm index 66081d7..17ebb1e 100644 --- a/lib/PCAP/Threaded.pm +++ b/lib/PCAP/Threaded.pm @@ -240,38 +240,42 @@ sub _create_script { my $script = "$stub.sh"; open my $SH, '>', $script or die "Cannot create $script: $!\n"; - print $SH qq{#!/bin/bash\nset -eux\n}; - print $SH join qq{\n}, @{$commands}; - print $SH qq{\n}; + print $SH qq{#!/bin/bash\nset -eux\n} or die "Write to $script failed"; + print $SH join qq{\n}, @{$commands}, q{} or die "Write to $script failed"; close $SH; undef $SH; - # sleep for random millisec max 1 sec. - my $millisec = rand 1_000_000; - $millisec = 100_000 if($millisec < 100_000); # min 0.1 sec + chmod $SCRIPT_OCT_MODE, $script or die "Failed to set executable flag on: $script"; - _file_complete($script, $millisec); - chmod $SCRIPT_OCT_MODE, $script; - usleep($millisec); + # sleep for random microsec max 1 sec. + my $microsec = int rand 1_000_000; + $microsec = 100_000 if($microsec < 200_000); # min 0.2 sec + _file_complete($script, $microsec); + usleep($microsec); return $script; } sub _file_complete { - my ($script, $millisec, $debug) = @_; + my ($script, $microsec, $debug) = @_; + $debug = 1; my $tries = 0; + while(! -e $script) { + $tries++; + croak "Failed to find script after 30 attempts ($microsec us delays): $script" if($tries >= 30); + usleep($microsec); + } + $tries = 0; while(1) { - croak "Failed to confirm script written after 30 attempts: $script" if($tries > 30); + $tries++; + croak "Failed to confirm write complete after 30 attempts ($microsec us delays): $script" if($tries >= 30); my ($stdout, $stderr, $exit) = capture { system([0,1], 'lsof', $script); }; if($exit > 1) { - croak $stderr; + croak sprintf "ERROR: lsof output\n\tSTDOUT: %s\n\tSTDERR: %s\n\tEXIT: %d\n", $stdout, $stderr, $exit; } printf STDERR "OUT : %s\nERR : %s\nEXIT: %s\n", $stdout,$stderr,$exit if($debug); - if($exit == 1) { - last if(-e $script); - } - warn $millisec if($debug); - usleep($millisec); - $tries++; + last if($exit == 1); + warn $microsec if($debug); + usleep($microsec); } return 1; } From bbf58df03b48d54aa0eca705149a2472e4b69c7a Mon Sep 17 00:00:00 2001 From: Keiran Raine Date: Fri, 27 Jan 2017 11:24:43 +0000 Subject: [PATCH 11/26] minor revision to reduce unneeded output --- lib/PCAP/Threaded.pm | 2 +- 1 file changed, 1 insertion(+), 1 deletion(-) diff --git a/lib/PCAP/Threaded.pm b/lib/PCAP/Threaded.pm index 17ebb1e..34381dc 100644 --- a/lib/PCAP/Threaded.pm +++ b/lib/PCAP/Threaded.pm @@ -272,7 +272,7 @@ sub _file_complete { if($exit > 1) { croak sprintf "ERROR: lsof output\n\tSTDOUT: %s\n\tSTDERR: %s\n\tEXIT: %d\n", $stdout, $stderr, $exit; } - printf STDERR "OUT : %s\nERR : %s\nEXIT: %s\n", $stdout,$stderr,$exit if($debug); + printf STDERR "OUT : %s\nERR : %s\nEXIT: %s\n", $stdout,$stderr,$exit if($debug && $exit == 0); last if($exit == 1); warn $microsec if($debug); usleep($microsec); From 244c8abc0b19eea6ae080135f2738a2656d24b70 Mon Sep 17 00:00:00 2001 From: Keiran Raine Date: Tue, 31 Jan 2017 09:35:18 +0000 Subject: [PATCH 12/26] Always output when evidence of an open file --- lib/PCAP/Threaded.pm | 6 ++---- 1 file changed, 2 insertions(+), 4 deletions(-) diff --git a/lib/PCAP/Threaded.pm b/lib/PCAP/Threaded.pm index 34381dc..81672d3 100644 --- a/lib/PCAP/Threaded.pm +++ b/lib/PCAP/Threaded.pm @@ -256,8 +256,7 @@ sub _create_script { } sub _file_complete { - my ($script, $microsec, $debug) = @_; - $debug = 1; + my ($script, $microsec) = @_; my $tries = 0; while(! -e $script) { $tries++; @@ -272,9 +271,8 @@ sub _file_complete { if($exit > 1) { croak sprintf "ERROR: lsof output\n\tSTDOUT: %s\n\tSTDERR: %s\n\tEXIT: %d\n", $stdout, $stderr, $exit; } - printf STDERR "OUT : %s\nERR : %s\nEXIT: %s\n", $stdout,$stderr,$exit if($debug && $exit == 0); + printf STDERR "OUT : %s\nERR : %s\nEXIT: %s\n", $stdout,$stderr,$exit if($exit == 0); last if($exit == 1); - warn $microsec if($debug); usleep($microsec); } return 1; From b1749c9506716b4961a0bbcc01e642068edadb37 Mon Sep 17 00:00:00 2001 From: Keiran Raine Date: Tue, 31 Jan 2017 09:37:41 +0000 Subject: [PATCH 13/26] extend dependancies --- INSTALL.md | 2 +- 1 file changed, 1 insertion(+), 1 deletion(-) diff --git a/INSTALL.md b/INSTALL.md index 00c7c33..a5526cf 100644 --- a/INSTALL.md +++ b/INSTALL.md @@ -29,7 +29,7 @@ bwa_mem.pl will only function when 0.7.x installed ``` apt-get update && \ apt-get -y install \ -build-essential zlib1g-dev libncurses5-dev libcurl4-gnutls-dev libssl-dev libexpat1-dev nettle-dev &&\ +build-essential zlib1g-dev libncurses5-dev libcurl4-gnutls-dev libssl-dev libexpat1-dev nettle-dev lsof libgoogle-perftools-dev &&\ apt-get clean ``` From 68507b38d0996a90dc0f076f6f3a78445e83195e Mon Sep 17 00:00:00 2001 From: Keiran Raine Date: Tue, 31 Jan 2017 09:43:50 +0000 Subject: [PATCH 14/26] Add deps --- .travis.yml | 2 ++ 1 file changed, 2 insertions(+) diff --git a/.travis.yml b/.travis.yml index b25ffcf..f21d3ac 100644 --- a/.travis.yml +++ b/.travis.yml @@ -21,6 +21,8 @@ addons: - unzip - libpng12-dev - libexpat1-dev + - libgoogle-perftools-dev + - lsof install: true From 683265674a670269aba54d0ce0d07b148530f9ef Mon Sep 17 00:00:00 2001 From: Keiran Raine Date: Tue, 31 Jan 2017 10:30:34 +0000 Subject: [PATCH 15/26] Resolves #139 --- bin/bwa_mem.pl | 1 + 1 file changed, 1 insertion(+) diff --git a/bin/bwa_mem.pl b/bin/bwa_mem.pl index 353f24d..40a71fe 100755 --- a/bin/bwa_mem.pl +++ b/bin/bwa_mem.pl @@ -161,6 +161,7 @@ sub setup { $_ = "$cwd/$_" unless($_ =~ m|^/|); push @{$opts{'raw_files'}}, $_; } + pod2usage(-msg => "\nERROR: No BAM/CRAM or FASTQ files have been defined.\n", -verbose => 1, -output => \*STDERR) if(scalar @{$opts{'raw_files'}} == 0); my $max_split = PCAP::Bwa::mem_prepare(\%opts); From fe83a12571e76dabdf9d892a7fc6a3de9a4d668b Mon Sep 17 00:00:00 2001 From: Keiran Raine Date: Tue, 31 Jan 2017 10:34:14 +0000 Subject: [PATCH 16/26] Update version and licence date, ad a trivial test --- lib/PCAP.pm | 4 ++-- t/pcap.t | 2 +- 2 files changed, 3 insertions(+), 3 deletions(-) diff --git a/lib/PCAP.pm b/lib/PCAP.pm index 8835cbe..d5f9015 100644 --- a/lib/PCAP.pm +++ b/lib/PCAP.pm @@ -27,12 +27,12 @@ use FindBin qw($Bin); use File::Which qw(which); # don't use autodie, only core perl in here -our $VERSION = '3.4.1'; +our $VERSION = '3.5.0'; our @EXPORT = qw($VERSION _which); const my $LICENSE => "################# -# PCAP version %s, Copyright (C) 2014-2016 ICGC/TCGA Pan-Cancer Analysis Project +# PCAP version %s, Copyright (C) 2014-2017 ICGC/TCGA Pan-Cancer Analysis Project # PCAP comes with ABSOLUTELY NO WARRANTY # See LICENSE for full details. #################"; diff --git a/t/pcap.t b/t/pcap.t index 174c1a7..a9f105c 100644 --- a/t/pcap.t +++ b/t/pcap.t @@ -10,9 +10,9 @@ subtest 'Initialisation checks' => sub { ok(PCAP::license(), 'License text retrieved'); +ok(PCAP->VERSION, 'Version retrieved'); is(PCAP::upgrade_path(), 'biobambam,samtools,bwa', 'Default program install when no previous version'); is(PCAP::upgrade_path('9.9.9'), 'biobambam,samtools,bwa', 'Default program install when unknown version installed'); done_testing(); - From e4ba46bcbc4e173570502616386724463335f4b2 Mon Sep 17 00:00:00 2001 From: Keiran Raine Date: Tue, 31 Jan 2017 14:08:34 +0000 Subject: [PATCH 17/26] Attempt to make file writes even more robust, optional force of sync via env variable --- lib/PCAP/Threaded.pm | 8 ++++++-- 1 file changed, 6 insertions(+), 2 deletions(-) diff --git a/lib/PCAP/Threaded.pm b/lib/PCAP/Threaded.pm index 81672d3..ed57ae0 100644 --- a/lib/PCAP/Threaded.pm +++ b/lib/PCAP/Threaded.pm @@ -239,11 +239,15 @@ sub _create_script { my ($commands, $stub) = @_; my $script = "$stub.sh"; - open my $SH, '>', $script or die "Cannot create $script: $!\n"; + + my $SH = IO::File->new($script, 'w'); + die "Cannot create $script: $!\n" unless(defined $SH); + #open my $SH, '>', $script or die "Cannot create $script: $!\n"; print $SH qq{#!/bin/bash\nset -eux\n} or die "Write to $script failed"; print $SH join qq{\n}, @{$commands}, q{} or die "Write to $script failed"; - close $SH; undef $SH; + autoflush STDOUT 1; + system('sync') if(exists $ENV{CGP_FORCE_SYNC}); chmod $SCRIPT_OCT_MODE, $script or die "Failed to set executable flag on: $script"; From 49e3cdae41f66b1f8e2fe5771b5f969a1b41761d Mon Sep 17 00:00:00 2001 From: Keiran Raine Date: Tue, 31 Jan 2017 14:08:47 +0000 Subject: [PATCH 18/26] Update HTS lib version --- setup.sh | 4 ++-- 1 file changed, 2 insertions(+), 2 deletions(-) diff --git a/setup.sh b/setup.sh index 7801985..33714a8 100755 --- a/setup.sh +++ b/setup.sh @@ -5,7 +5,7 @@ SOURCE_BWA="https://github.com/lh3/bwa/archive/v0.7.15.tar.gz" SOURCE_SAMTOOLS="https://github.com/samtools/samtools/releases/download/1.3.1/samtools-1.3.1.tar.bz2" # for bamstats and Bio::DB::HTS -SOURCE_HTSLIB="https://github.com/samtools/htslib/releases/download/1.3.1/htslib-1.3.1.tar.bz2" +SOURCE_HTSLIB="https://github.com/samtools/htslib/releases/download/1.3.2/htslib-1.3.2.tar.bz2" # Bio::DB::HTS SOURCE_BIOBDHTS="https://github.com/Ensembl/Bio-HTS/archive/2.3.tar.gz" @@ -285,7 +285,6 @@ else env HTSLIB=$SETUP_DIR/htslib make -C c -j$CPU prefix=$INST_PATH cp bin/bam_stats $INST_PATH/bin/. cp bin/reheadSQ $INST_PATH/bin/. - cp bin/diff_bams $INST_PATH/bin/. touch $SETUP_DIR/bam_stats.success make -C c clean fi @@ -318,3 +317,4 @@ echo " $PERLROOT" echo exit 0 + From 0101a5850e717435009ed5fd1d0301eb8715baa7 Mon Sep 17 00:00:00 2001 From: Keiran Raine Date: Tue, 31 Jan 2017 14:28:02 +0000 Subject: [PATCH 19/26] More optional file checks --- lib/PCAP/Threaded.pm | 5 ++++- 1 file changed, 4 insertions(+), 1 deletion(-) diff --git a/lib/PCAP/Threaded.pm b/lib/PCAP/Threaded.pm index ed57ae0..a2689bd 100644 --- a/lib/PCAP/Threaded.pm +++ b/lib/PCAP/Threaded.pm @@ -240,6 +240,9 @@ sub _create_script { my $script = "$stub.sh"; + if(exists $ENV{PCAP_THREADED_NO_SCRIPT} && -e $script) { + die "ERROR: Script already present, delete to proceed: $script"; + } my $SH = IO::File->new($script, 'w'); die "Cannot create $script: $!\n" unless(defined $SH); #open my $SH, '>', $script or die "Cannot create $script: $!\n"; @@ -247,7 +250,7 @@ sub _create_script { print $SH join qq{\n}, @{$commands}, q{} or die "Write to $script failed"; undef $SH; autoflush STDOUT 1; - system('sync') if(exists $ENV{CGP_FORCE_SYNC}); + system('sync') if(exists $ENV{PCAP_THREADED_FORCE_SYNC}); chmod $SCRIPT_OCT_MODE, $script or die "Failed to set executable flag on: $script"; From f0408e7c52946c9ba72676b0b0c20a4d0ce3d369 Mon Sep 17 00:00:00 2001 From: Keiran Raine Date: Wed, 1 Feb 2017 08:49:29 +0000 Subject: [PATCH 20/26] Remove comment --- lib/PCAP/Threaded.pm | 1 - 1 file changed, 1 deletion(-) diff --git a/lib/PCAP/Threaded.pm b/lib/PCAP/Threaded.pm index a2689bd..d43f663 100644 --- a/lib/PCAP/Threaded.pm +++ b/lib/PCAP/Threaded.pm @@ -245,7 +245,6 @@ sub _create_script { } my $SH = IO::File->new($script, 'w'); die "Cannot create $script: $!\n" unless(defined $SH); - #open my $SH, '>', $script or die "Cannot create $script: $!\n"; print $SH qq{#!/bin/bash\nset -eux\n} or die "Write to $script failed"; print $SH join qq{\n}, @{$commands}, q{} or die "Write to $script failed"; undef $SH; From 2bc73ae9785a56e5e10648d952de31be388f76ea Mon Sep 17 00:00:00 2001 From: Keiran Raine Date: Wed, 1 Feb 2017 09:20:22 +0000 Subject: [PATCH 21/26] cleanup some null chars --- setup.sh | 1 - 1 file changed, 1 deletion(-) diff --git a/setup.sh b/setup.sh index 33714a8..f5d9ae6 100755 --- a/setup.sh +++ b/setup.sh @@ -317,4 +317,3 @@ echo " $PERLROOT" echo exit 0 - From 21ea8770c7bb64d6fc1e23e56f6e77312c73006e Mon Sep 17 00:00:00 2001 From: Keiran Raine Date: Wed, 1 Feb 2017 09:30:37 +0000 Subject: [PATCH 22/26] ensure diff_bams is deployed --- setup.sh | 1 + 1 file changed, 1 insertion(+) diff --git a/setup.sh b/setup.sh index f5d9ae6..d4d179f 100755 --- a/setup.sh +++ b/setup.sh @@ -285,6 +285,7 @@ else env HTSLIB=$SETUP_DIR/htslib make -C c -j$CPU prefix=$INST_PATH cp bin/bam_stats $INST_PATH/bin/. cp bin/reheadSQ $INST_PATH/bin/. + cp bin/diff_bams $INST_PATH/bin/. touch $SETUP_DIR/bam_stats.success make -C c clean fi From 73a92e2475c05c5e3af9ba680cfa288dec50012d Mon Sep 17 00:00:00 2001 From: Keiran Raine Date: Wed, 1 Feb 2017 09:30:49 +0000 Subject: [PATCH 23/26] Update release information --- CHANGES.md | 12 ++++++++++++ 1 file changed, 12 insertions(+) diff --git a/CHANGES.md b/CHANGES.md index 57bbcf8..3df33cc 100644 --- a/CHANGES.md +++ b/CHANGES.md @@ -1,3 +1,15 @@ +### 3.5.0 +* Adds RNA downloads to PanCancer download tool `gnos_pull.pl` +* Hardening of external process handling in `PCAP::Threaded` +* Adds C version of `diff_bams` + +### 3.4.0 +* Significant speed up of BAM generation under `bwa_mem.pl` by using separate process to do compression of mark duplicate output and streaming BAS generation. Not possible to do this to CRAM in same way. + +### 3.3.4 +* Reduce disk usage when running `bwa_mem.pl` +* Improve throughput via slightly unintuitive use of additional pipes + ### 3.3.0 * adds `map_threads|mt` option to `bwa_mem.pl` to allow more control of parallel processing in one shot submission. * adds `bwa_pl|l` option to `bwa_mem.pl` to allow preload of different malloc libraries. From 5c83d73c9f105268ef291f4f5d5c6a5ffff7819b Mon Sep 17 00:00:00 2001 From: Keiran Raine Date: Wed, 1 Feb 2017 09:34:56 +0000 Subject: [PATCH 24/26] Additional checks of critical items --- .travis.yml | 4 ++++ 1 file changed, 4 insertions(+) diff --git a/.travis.yml b/.travis.yml index f21d3ac..641c7a1 100644 --- a/.travis.yml +++ b/.travis.yml @@ -40,3 +40,7 @@ before_script: script: - ./setup.sh ~/wtsi-opt - ~/wtsi-opt/bin/samtools view # dump usage to show intact + - ~/wtsi-opt/bin/diff_bams -h + - ~/wtsi-opt/bin/reheadSQ -h + - ~/wtsi-opt/bin/bam_stats -h + - ~/wtsi-opt/bin/bwa_mem.pl -h From c2b7bdde7529e22af5e79d834fbf9f4dd3ea67e0 Mon Sep 17 00:00:00 2001 From: Keiran Raine Date: Wed, 1 Feb 2017 09:35:17 +0000 Subject: [PATCH 25/26] release files --- MANIFEST | 129 +--------------------------------------------------- docs.tar.gz | Bin 173087 -> 174684 bytes 2 files changed, 1 insertion(+), 128 deletions(-) diff --git a/MANIFEST b/MANIFEST index 9976522..ae492ba 100644 --- a/MANIFEST +++ b/MANIFEST @@ -1,29 +1,4 @@ .travis.yml -bam_out/logs_bwamem_wibble/PCAP_Bam_merge_and_mark_dup.0.err -bam_out/logs_bwamem_wibble/PCAP_Bam_merge_and_mark_dup.0.out -bam_out/logs_bwamem_wibble/PCAP_Bwa_bwa_mem.1.err -bam_out/logs_bwamem_wibble/PCAP_Bwa_bwa_mem.1.out -bam_out/logs_bwamem_wibble/PCAP_Bwa_bwa_mem.2.err -bam_out/logs_bwamem_wibble/PCAP_Bwa_bwa_mem.2.out -bam_out/logs_bwamem_wibble/PCAP_Bwa_bwa_mem.3.err -bam_out/logs_bwamem_wibble/PCAP_Bwa_bwa_mem.3.out -bam_out/logs_bwamem_wibble/PCAP_Bwa_bwa_mem.4.err -bam_out/logs_bwamem_wibble/PCAP_Bwa_bwa_mem.4.out -bam_out/logs_bwamem_wibble/PCAP_Bwa_bwa_mem.5.err -bam_out/logs_bwamem_wibble/PCAP_Bwa_bwa_mem.5.out -bam_out/logs_bwamem_wibble/PCAP_Bwa_bwa_mem.6.err -bam_out/logs_bwamem_wibble/PCAP_Bwa_bwa_mem.6.out -bam_out/logs_bwamem_wibble/PCAP_Bwa_bwa_mem.7.err -bam_out/logs_bwamem_wibble/PCAP_Bwa_bwa_mem.7.out -bam_out/logs_bwamem_wibble/PCAP_Bwa_bwa_mem.8.err -bam_out/logs_bwamem_wibble/PCAP_Bwa_bwa_mem.8.out -bam_out/logs_bwamem_wibble/PCAP_Bwa_split_in.1.err -bam_out/logs_bwamem_wibble/PCAP_Bwa_split_in.1.out -bam_out/wibble.bam -bam_out/wibble.bam.bai -bam_out/wibble.bam.bas -bam_out/wibble.bam.md5 -bam_out/wibble.bam.met bin/bam_stats.pl bin/bam_to_sra_sub.pl bin/bamToBw.pl @@ -49,6 +24,7 @@ c/c_tests/minunit.h c/c_tests/runtests.sh c/c_tests/tests_log c/dbg.h +c/diff_bams.c c/khash.h c/reheadSQ.c CHANGES.md @@ -56,109 +32,6 @@ dists/patch/Bio-BigFile_build.patch dists/snappy-1.1.2.tar.gz docs.tar.gz examples/gnos_pull.ini -fastq/1/10658_1.fq.gz -fastq/1/10659_1.fq.gz -fastq/1/10660_1.fq.gz -fastq/1/10661_1.fq.gz -fastq/1/10662_1.fq.gz -fastq/1/10663_1.fq.gz -fastq/1/10664_1.fq.gz -fastq/1/10665_1.fq.gz -fastq/2/10658_2.fq.gz -fastq/2/10659_2.fq.gz -fastq/2/10660_2.fq.gz -fastq/2/10661_2.fq.gz -fastq/2/10662_2.fq.gz -fastq/2/10663_2.fq.gz -fastq/2/10664_2.fq.gz -fastq/2/10665_2.fq.gz -fastq_i/10658_i.fq.gz -fastq_i/10659_i.fq.gz -fastq_i/10660_i.fq.gz -fastq_i/10661_i.fq.gz -fastq_i/10662_i.fq.gz -fastq_i/10663_i.fq.gz -fastq_i/10664_i.fq.gz -fastq_i/10665_i.fq.gz -fastq_i_out/logs_bwamem_wibble/PCAP_Bam_merge_and_mark_dup.0.err -fastq_i_out/logs_bwamem_wibble/PCAP_Bam_merge_and_mark_dup.0.out -fastq_i_out/logs_bwamem_wibble/PCAP_Bwa_bwa_mem.1.err -fastq_i_out/logs_bwamem_wibble/PCAP_Bwa_bwa_mem.1.out -fastq_i_out/logs_bwamem_wibble/PCAP_Bwa_bwa_mem.2.err -fastq_i_out/logs_bwamem_wibble/PCAP_Bwa_bwa_mem.2.out -fastq_i_out/logs_bwamem_wibble/PCAP_Bwa_bwa_mem.3.err -fastq_i_out/logs_bwamem_wibble/PCAP_Bwa_bwa_mem.3.out -fastq_i_out/logs_bwamem_wibble/PCAP_Bwa_bwa_mem.4.err -fastq_i_out/logs_bwamem_wibble/PCAP_Bwa_bwa_mem.4.out -fastq_i_out/logs_bwamem_wibble/PCAP_Bwa_bwa_mem.5.err -fastq_i_out/logs_bwamem_wibble/PCAP_Bwa_bwa_mem.5.out -fastq_i_out/logs_bwamem_wibble/PCAP_Bwa_bwa_mem.6.err -fastq_i_out/logs_bwamem_wibble/PCAP_Bwa_bwa_mem.6.out -fastq_i_out/logs_bwamem_wibble/PCAP_Bwa_bwa_mem.7.err -fastq_i_out/logs_bwamem_wibble/PCAP_Bwa_bwa_mem.7.out -fastq_i_out/logs_bwamem_wibble/PCAP_Bwa_bwa_mem.8.err -fastq_i_out/logs_bwamem_wibble/PCAP_Bwa_bwa_mem.8.out -fastq_i_out/logs_bwamem_wibble/PCAP_Bwa_split_in.1.err -fastq_i_out/logs_bwamem_wibble/PCAP_Bwa_split_in.1.out -fastq_i_out/logs_bwamem_wibble/PCAP_Bwa_split_in.2.err -fastq_i_out/logs_bwamem_wibble/PCAP_Bwa_split_in.2.out -fastq_i_out/logs_bwamem_wibble/PCAP_Bwa_split_in.3.err -fastq_i_out/logs_bwamem_wibble/PCAP_Bwa_split_in.3.out -fastq_i_out/logs_bwamem_wibble/PCAP_Bwa_split_in.4.err -fastq_i_out/logs_bwamem_wibble/PCAP_Bwa_split_in.4.out -fastq_i_out/logs_bwamem_wibble/PCAP_Bwa_split_in.5.err -fastq_i_out/logs_bwamem_wibble/PCAP_Bwa_split_in.5.out -fastq_i_out/logs_bwamem_wibble/PCAP_Bwa_split_in.6.err -fastq_i_out/logs_bwamem_wibble/PCAP_Bwa_split_in.6.out -fastq_i_out/logs_bwamem_wibble/PCAP_Bwa_split_in.7.err -fastq_i_out/logs_bwamem_wibble/PCAP_Bwa_split_in.7.out -fastq_i_out/logs_bwamem_wibble/PCAP_Bwa_split_in.8.err -fastq_i_out/logs_bwamem_wibble/PCAP_Bwa_split_in.8.out -fastq_i_out/wibble.bam -fastq_i_out/wibble.bam.bai -fastq_i_out/wibble.bam.bas -fastq_i_out/wibble.bam.md5 -fastq_i_out/wibble.bam.met -fastq_out/logs_bwamem_wibble/PCAP_Bam_merge_and_mark_dup.0.err -fastq_out/logs_bwamem_wibble/PCAP_Bam_merge_and_mark_dup.0.out -fastq_out/logs_bwamem_wibble/PCAP_Bwa_bwa_mem.1.err -fastq_out/logs_bwamem_wibble/PCAP_Bwa_bwa_mem.1.out -fastq_out/logs_bwamem_wibble/PCAP_Bwa_bwa_mem.2.err -fastq_out/logs_bwamem_wibble/PCAP_Bwa_bwa_mem.2.out -fastq_out/logs_bwamem_wibble/PCAP_Bwa_bwa_mem.3.err -fastq_out/logs_bwamem_wibble/PCAP_Bwa_bwa_mem.3.out -fastq_out/logs_bwamem_wibble/PCAP_Bwa_bwa_mem.4.err -fastq_out/logs_bwamem_wibble/PCAP_Bwa_bwa_mem.4.out -fastq_out/logs_bwamem_wibble/PCAP_Bwa_bwa_mem.5.err -fastq_out/logs_bwamem_wibble/PCAP_Bwa_bwa_mem.5.out -fastq_out/logs_bwamem_wibble/PCAP_Bwa_bwa_mem.6.err -fastq_out/logs_bwamem_wibble/PCAP_Bwa_bwa_mem.6.out 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