-#!/usr/bin/perl
+#!/usr/bin/env perl
use Getopt::Long;
use Pod::Usage;
-use FindBin;
-use lib $FindBin::Bin;
-use strict;
+use FindBin;
+use lib $FindBin::RealBin;
use rsem_perl_utils qw(runCommand collectResults showVersionInfo);
+use Env qw(@PATH);
+@PATH = ($FindBin::RealBin, "$FindBin::RealBin/sam", @PATH);
+
+use strict;
+
#const
my $BURNIN = 200;
my $NCV = 1000;
my $sampling = 0;
my $calcPME = 0;
my $calcCI = 0;
-my $var_opt = 0; # temporarily, only for internal use
my $quiet = 0;
my $help = 0;
my $bowtie2_k = 200;
my $bowtie2_sensitivity_level = "sensitive"; # must be one of "very_fast", "fast", "sensitive", "very_sensitive"
+my $seed = "NULL";
+
my $version = 0;
my $mTime = 0;
"output-genome-bam" => \$genGenomeBamF,
"sampling-for-bam" => \$sampling,
"calc-pme" => \$calcPME,
- "var" => \$var_opt,
"calc-ci" => \$calcCI,
"ci-memory=i" => \$NMB,
"samtools-sort-mem=s" => \$SortMem,
+ "seed=i" => \$seed,
"time" => \$mTime,
"version" => \$version,
"q|quiet" => \$quiet,
"h|help" => \$help) or pod2usage(-exitval => 2, -verbose => 2);
-my $dir = "$FindBin::Bin/";
-
pod2usage(-verbose => 2) if ($help == 1);
-&showVersionInfo($dir) if ($version == 1);
+&showVersionInfo($FindBin::RealBin) if ($version == 1);
#check parameters and options
pod2usage(-msg => "Seed length should be at least 5!\n", -exitval => 2, -verbose => 2) if ($L < 5);
pod2usage(-msg => "--sampling-for-bam cannot be specified if --no-bam-output is specified!\n", -exitval => 2, -verbose => 2) if ($sampling && !$genBamF);
pod2usage(-msg => "--output-genome-bam cannot be specified if --no-bam-output is specified!\n", -exitval => 2, -verbose => 2) if ($genGenomeBamF && !$genBamF);
+pod2usage(-msg => "The seed for random number generator must be a non-negative 32bit integer!\n", -exitval => 2, -verbose => 2) if (($seed ne "NULL") && ($seed < 0 || $seed > 0xffffffff));
if ($L < 25) { print "Warning: the seed length set is less than 25! This is only allowed if the references are not added poly(A) tails.\n"; }
}
# pipe to samtools to generate a BAM file
- $command .= " | $dir\sam/samtools view -S -b -o $imdName.bam -";
+ $command .= " | samtools view -S -b -o $imdName.bam -";
}
else {
$command = $bowtie2_path."bowtie2";
}
# pipe to samtools to generate a BAM file
- $command .= " | $dir\sam/samtools view -S -b -o $imdName.bam -";
+ $command .= " | samtools view -S -b -o $imdName.bam -";
}
if ($mTime) { $time_start = time(); }
if ($mTime) { $time_start = time(); }
-$command = $dir."rsem-parse-alignments $refName $imdName $statName";
+$command = "rsem-parse-alignments $refName $imdName $statName";
my $samInpType;
if ($is_sam) { $samInpType = "s"; }
&runCommand($command);
-$command = $dir."rsem-build-read-index $gap";
+$command = "rsem-build-read-index $gap";
if ($read_type == 0) { $command .= " 0 $quiet $imdName\_alignable.fa"; }
elsif ($read_type == 1) { $command .= " 1 $quiet $imdName\_alignable.fq"; }
elsif ($read_type == 2) { $command .= " 0 $quiet $imdName\_alignable_1.fa $imdName\_alignable_2.fa"; }
print OUTPUT "$L\n";
close(OUTPUT);
-$command = $dir."rsem-run-em $refName $read_type $sampleName $imdName $statName -p $nThreads";
+my @seeds = ();
+if ($seed ne "NULL") {
+ srand($seed);
+ for (my $i = 0; $i < 3; $i++) {
+ push(@seeds, int(rand(1 << 32)));
+ }
+}
+
+$command = "rsem-run-em $refName $read_type $sampleName $imdName $statName -p $nThreads";
if ($genBamF) {
$command .= " -b $samInpType $inpF";
if ($fn_list ne "") { $command .= " 1 $fn_list"; }
else { $command .= " 0"; }
if ($sampling) { $command .= " --sampling"; }
+ if ($seed ne "NULL") { $command .= " --seed $seeds[0]"; }
}
-if ($calcPME || $var_opt || $calcCI) { $command .= " --gibbs-out"; }
+if ($calcPME || $calcCI) { $command .= " --gibbs-out"; }
if ($quiet) { $command .= " -q"; }
&runCommand($command);
}
if ($genBamF) {
- $command = $dir."sam/samtools sort -@ $nThreads -m $SortMem $sampleName.transcript.bam $sampleName.transcript.sorted";
+ $command = "samtools sort -@ $nThreads -m $SortMem $sampleName.transcript.bam $sampleName.transcript.sorted";
&runCommand($command);
- $command = $dir."sam/samtools index $sampleName.transcript.sorted.bam";
+ $command = "samtools index $sampleName.transcript.sorted.bam";
&runCommand($command);
if ($genGenomeBamF) {
- $command = $dir."rsem-tbam2gbam $refName $sampleName.transcript.bam $sampleName.genome.bam";
+ $command = "rsem-tbam2gbam $refName $sampleName.transcript.bam $sampleName.genome.bam";
&runCommand($command);
- $command = $dir."sam/samtools sort -@ $nThreads -m $SortMem $sampleName.genome.bam $sampleName.genome.sorted";
+ $command = "samtools sort -@ $nThreads -m $SortMem $sampleName.genome.bam $sampleName.genome.sorted";
&runCommand($command);
- $command = $dir."sam/samtools index $sampleName.genome.sorted.bam";
+ $command = "samtools index $sampleName.genome.sorted.bam";
&runCommand($command);
}
}
if ($mTime) { $time_start = time(); }
-if ($calcPME || $var_opt || $calcCI ) {
- $command = $dir."rsem-run-gibbs $refName $imdName $statName $BURNIN $NCV $SAMPLEGAP";
+if ($calcPME || $calcCI ) {
+ $command = "rsem-run-gibbs $refName $imdName $statName $BURNIN $NCV $SAMPLEGAP";
$command .= " -p $nThreads";
- if ($var_opt) { $command .= " --var"; }
+ if ($seed ne "NULL") { $command .= " --seed $seeds[1]"; }
if ($quiet) { $command .= " -q"; }
&runCommand($command);
}
}
if ($calcCI) {
- $command = $dir."rsem-calculate-credibility-intervals $refName $imdName $statName $CONFIDENCE $NCV $NSPC $NMB";
+ $command = "rsem-calculate-credibility-intervals $refName $imdName $statName $CONFIDENCE $NCV $NSPC $NMB";
$command .= " -p $nThreads";
+ if ($seed ne "NULL") { $command .= " --seed $seeds[2]"; }
if ($quiet) { $command .= " -q"; }
&runCommand($command);
When RSEM generates a BAM file, instead of outputing all alignments a read has with their posterior probabilities, one alignment is sampled according to the posterior probabilities. The sampling procedure includes the alignment to the "noise" transcript, which does not appear in the BAM file. Only the sampled alignment has a weight of 1. All other alignments have weight 0. If the "noise" transcript is sampled, all alignments appeared in the BAM file should have weight 0. (Default: off)
+=item B<--seed> <uint32>
+
+Set the seed for the random number generators used in calculating posterior mean estimates and credibility intervals. The seed must be a non-negative 32 bit interger. (Default: off)
+
=item B<--calc-pme>
Run RSEM's collapsed Gibbs sampler to calculate posterior mean estimates. (Default: off)
contains column names separated by the tab character. The format of
each line in the rest of this file is:
-transcript_id gene_id length effective_length expected_count TPM FPKM IsoPct [pme_expected_count pme_TPM pme_FPKM IsoPct_from_pme_TPM TPM_ci_lower_bound TPM_ci_upper_bound FPKM_ci_lower_bound FPKM_ci_upper_bound]
+transcript_id gene_id length effective_length expected_count TPM FPKM IsoPct [posterior_mean_count posterior_standard_deviation_of_count pme_TPM pme_FPKM IsoPct_from_pme_TPM TPM_ci_lower_bound TPM_ci_upper_bound FPKM_ci_lower_bound FPKM_ci_upper_bound]
Fields are separated by the tab character. Fields within "[]" are
optional. They will not be presented if neither '--calc-pme' nor
gene has only one isoform or the gene information is not provided,
this field will be set to 100.
-'pme_expected_count', 'pme_TPM', 'pme_FPKM' are posterior mean
-estimates calculated by RSEM's Gibbs sampler. 'IsoPct_from_pme_TPM' is
-the isoform percentage calculated from 'pme_TPM' values.
+'posterior_mean_count', 'pme_TPM', 'pme_FPKM' are posterior mean
+estimates calculated by RSEM's Gibbs
+sampler. 'posterior_standard_deviation_of_count' is the posterior
+standard deviation of counts. 'IsoPct_from_pme_TPM' is the isoform
+percentage calculated from 'pme_TPM' values.
'TPM_ci_lower_bound', 'TPM_ci_upper_bound', 'FPKM_ci_lower_bound' and
'FPKM_ci_upper_bound' are lower(l) and upper(u) bounds of 95%
contains column names separated by the tab character. The format of
each line in the rest of this file is:
-gene_id transcript_id(s) length effective_length expected_count TPM FPKM [pme_expected_count pme_TPM pme_FPKM TPM_ci_lower_bound TPM_ci_upper_bound FPKM_ci_lower_bound FPKM_ci_upper_bound]
+gene_id transcript_id(s) length effective_length expected_count TPM FPKM [posterior_mean_count posterior_standard_deviation_of_count pme_TPM pme_FPKM TPM_ci_lower_bound TPM_ci_upper_bound FPKM_ci_lower_bound FPKM_ci_upper_bound]
Fields are separated by the tab character. Fields within "[]" are
optional. They will not be presented if neither '--calc-pme' nor
contains column names separated by the tab character. The format of
each line in the rest of this file is:
-allele_id transcript_id gene_id length effective_length expected_count TPM FPKM AlleleIsoPct AlleleGenePct [pme_expected_count pme_TPM pme_FPKM AlleleIsoPct_from_pme_TPM AlleleGenePct_from_pme_TPM TPM_ci_lower_bound TPM_ci_upper_bound FPKM_ci_lower_bound FPKM_ci_upper_bound]
+allele_id transcript_id gene_id length effective_length expected_count TPM FPKM AlleleIsoPct AlleleGenePct [posterior_mean_count posterior_standard_deviation_of_count pme_TPM pme_FPKM AlleleIsoPct_from_pme_TPM AlleleGenePct_from_pme_TPM TPM_ci_lower_bound TPM_ci_upper_bound FPKM_ci_lower_bound FPKM_ci_upper_bound]
Fields are separated by the tab character. Fields within "[]" are
optional. They will not be presented if neither '--calc-pme' nor
projects / rsem.git / blobdiff