Provided by: rsem_1.2.31+dfsg-1_amd64 bug

NAME

       rsem-calculate-expression

PURPOSE

       Estimate gene and isoform expression from RNA-Seq data.

SYNOPSIS

        rsem-calculate-expression [options] upstream_read_file(s) reference_name sample_name
        rsem-calculate-expression [options] --paired-end upstream_read_file(s) downstream_read_file(s) reference_name sample_name
        rsem-calculate-expression [options] --alignments [--paired-end] input reference_name sample_name

ARGUMENTS

       upstream_read_files(s)
           Comma-separated  list of files containing single-end reads or upstream reads for paired-end data.  By
           default, these files are assumed to be in FASTQ format.  If the --no-qualities option  is  specified,
           then FASTA format is expected.

       downstream_read_file(s)
           Comma-separated  list  of files containing downstream reads which are paired with the upstream reads.
           By default, these files are assumed  to  be  in  FASTQ  format.   If  the  --no-qualities  option  is
           specified, then FASTA format is expected.

       input
           SAM/BAM/CRAM  formatted  input  file.   If  "-"  is  specified for the filename, the input is instead
           assumed to come from standard input. RSEM requires all alignments of the same  read  group  together.
           For  paired-end  reads,  RSEM  also requires the two mates of any alignment be adjacent. In addition,
           RSEM does not allow the SEQ and QUAL fields to be empty. See Description  section  for  how  to  make
           input file obey RSEM's requirements.

       reference_name
           The  name  of  the  reference  used.   The  user  must  have  run  'rsem-prepare-reference' with this
           reference_name before running this program.

       sample_name
           The  name  of  the  sample  analyzed.  All  output  files  are   prefixed   by   this   name   (e.g.,
           sample_name.genes.results)

BASIC OPTIONS

       --paired-end
           Input reads are paired-end reads. (Default: off)

       --no-qualities
           Input reads do not contain quality scores. (Default: off)

       --strand-specific
           The  RNA-Seq  protocol  used to generate the reads is strand specific, i.e., all (upstream) reads are
           derived from the forward strand.  This option is equivalent to --forward-prob=1.0.  With this  option
           set,  if  RSEM  runs  the  Bowtie/Bowtie 2 aligner, the '--norc' Bowtie/Bowtie 2 option will be used,
           which disables alignment to the reverse strand of transcripts.  (Default: off)

       -p/--num-threads <int>
           Number of threads to use. Both Bowtie/Bowtie2, expression estimation and  'samtools  sort'  will  use
           this many threads. (Default: 1)

       --alignments
           Input  file  contains  alignments  in  SAM/BAM/CRAM  format. The exact file format will be determined
           automatically. (Default: off)

       --fai <file>
           If the header section of input alignment file does not contain reference sequence  information,  this
           option should be turned on. <file> is a FAI format file containing each reference sequence's name and
           length. Please refer to the SAM official website for the details of FAI format. (Default: off)

       --bowtie2
           Use  Bowtie 2 instead of Bowtie to align reads. Since currently RSEM does not handle indel, local and
           discordant alignments, the Bowtie2 parameters are  set  in  a  way  to  avoid  those  alignments.  In
           particular,  we  use  options  '--sensitive  --dpad  0  --gbar  99999999  --mp 1,1 --np 1 --score-min
           L,0,-0.1' by default. The last parameter  of  '--score-min',  '-0.1',  is  the  negative  of  maximum
           mismatch  rate. This rate can be set by option '--bowtie2-mismatch-rate'. If reads are paired-end, we
           additionally use options '--no-mixed' and '--no-discordant'. (Default: off)

       --star
           Use STAR to align reads.  Alignment  parameters  are  from  ENCODE3's  STAR-RSEM  pipeline.  To  save
           computational  time  and memory resources, STAR's Output BAM file is unsorted. It is stored in RSEM's
           temporary directory with name as 'sample_name.bam'. Each STAR job will have its own private  copy  of
           the genome in memory. (Default: off)

       --append-names
           If  gene_name/transcript_name  is available, append it to the end of gene_id/transcript_id (separated
           by '_') in files 'sample_name.isoforms.results' and 'sample_name.genes.results'. (Default: off)

       --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 integer. (Default: off)

       --single-cell-prior
           By default, RSEM uses Dirichlet(1) as the prior to calculate posterior mean estimates and credibility
           intervals.  However,  much less genes are expressed in single cell RNA-Seq data. Thus, if you want to
           compute posterior mean estimates and/or credibility intervals and you have single-cell RNA-Seq  data,
           you  are  recommended  to  turn  on this option. Then RSEM will use Dirichlet(0.1) as the prior which
           encourage the sparsity of the expression levels. (Default: off)

       --calc-pme
           Run RSEM's collapsed Gibbs sampler to calculate posterior mean estimates. (Default: off)

       --calc-ci
           Calculate 95% credibility intervals and posterior  mean  estimates.  The  credibility  level  can  be
           changed by setting '--ci-credibility-level'. (Default: off)

       -q/--quiet
           Suppress the output of logging information. (Default: off)

       -h/--help
           Show help information.

       --version
           Show version information.

OUTPUT OPTIONS

       --sort-bam-by-read-name
           Sort  BAM  file aligned under transcript coordidate by read name. Setting this option on will produce
           deterministic maximum likelihood estimations from independent runs. Note that sorting will take  long
           time and lots of memory. (Default: off)

       --no-bam-output
           Do not output any BAM file. (Default: off)

       --sampling-for-bam
           When  RSEM generates a BAM file, instead of outputting 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)

       --output-genome-bam
           Generate  a  BAM  file,  'sample_name.genome.bam',  with alignments mapped to genomic coordinates and
           annotated with their posterior probabilities. In addition, RSEM will call samtools (included in  RSEM
           package)    to    sort    and    index    the    bam    file.   'sample_name.genome.sorted.bam'   and
           'sample_name.genome.sorted.bam.bai' will be generated. (Default: off)

       --sort-bam-by-coordinate
           Sort RSEM generated transcript and genome BAM files by  coordinates  and  build  associated  indices.
           (Default: off)

       --sort-bam-memory-per-thread <string>
           Set the maximum memory per thread that can be used by 'samtools sort'. <string> represents the memory
           and  accepts  suffices  'K/M/G'.  RSEM will pass <string> to the '-m' option of 'samtools sort'. Note
           that the default used here is different from the default used by samtools. (Default: 1G)

ALIGNER OPTIONS

       --seed-length <int>
           Seed length used by the read aligner.  Providing the correct value is important  for  RSEM.  If  RSEM
           runs Bowtie, it uses this value for Bowtie's seed length parameter. Any read with its or at least one
           of  its  mates' (for paired-end reads) length less than this value will be ignored. If the references
           are not added poly(A) tails, the minimum allowed value is 5, otherwise, the minimum allowed value  is
           25. Note that this script will only check if the value >= 5 and give a warning message if the value <
           25 but >= 5. (Default: 25)

       --phred33-quals
           Input quality scores are encoded as Phred+33. (Default: on)

       --phred64-quals
           Input quality scores are encoded as Phred+64 (default for GA Pipeline ver. >= 1.3). (Default: off)

       --solexa-quals
           Input quality scores are solexa encoded (from GA Pipeline ver. < 1.3). (Default: off)

       --bowtie-path <path>
           The  path to the Bowtie executables. (Default: the path to the Bowtie executables is assumed to be in
           the user's PATH environment variable)

       --bowtie-n <int>
           (Bowtie parameter) max # of mismatches in the seed. (Range: 0-3, Default: 2)

       --bowtie-e <int>
           (Bowtie parameter) max sum of mismatch quality scores across the alignment. (Default: 99999999)

       --bowtie-m <int>
           (Bowtie parameter) suppress all alignments for a read if > <int> valid  alignments  exist.  (Default:
           200)

       --bowtie-chunkmbs <int>
           (Bowtie  parameter)  memory allocated for best first alignment calculation (Default: 0 - use Bowtie's
           default)

       --bowtie2-path <path>
           (Bowtie 2 parameter) The path to the Bowtie 2  executables.  (Default:  the  path  to  the  Bowtie  2
           executables is assumed to be in the user's PATH environment variable)

       --bowtie2-mismatch-rate <double>
           (Bowtie 2 parameter) The maximum mismatch rate allowed. (Default: 0.1)

       --bowtie2-k <int>
           (Bowtie 2 parameter) Find up to <int> alignments per read. (Default: 200)

       --bowtie2-sensitivity-level <string>
           (Bowtie  2  parameter)  Set  Bowtie 2's preset options in --end-to-end mode. This option controls how
           hard Bowtie 2 tries to find alignments. <string> must be one of "very_fast", "fast", "sensitive"  and
           "very_sensitive". The four candidates correspond to Bowtie 2's "--very-fast", "--fast", "--sensitive"
           and "--very-sensitive" options. (Default: "sensitive" - use Bowtie 2's default)

       --star-path <path>
           The  path to STAR's executable. (Default: the path to STAR executable is assumed to be in user's PATH
           environment variable)

       --star-gzipped-read-file
           (STAR parameter) Input read file(s) is compressed by gzip. (Default: off)

       --star-bzipped-read-file
           (STAR parameter) Input read file(s) is compressed by bzip2. (Default: off)

       --star-output-genome-bam
           (STAR  parameter)  Save  the  BAM  file   from   STAR   alignment   under   genomic   coordinate   to
           'sample_name.STAR.genome.bam'. This file is NOT sorted by genomic coordinate. In this file, according
           to STAR's manual, 'paired ends of an alignment are always adjacent, and multiple alignments of a read
           are adjacent as well'. (Default: off)

ADVANCED OPTIONS

       --tag <string>
           The  name  of  the  optional  field  used in the SAM input for identifying a read with too many valid
           alignments. The field should have the format <tagName>:i:<value>,  where  a  <value>  bigger  than  0
           indicates a read with too many alignments. (Default: "")

       --forward-prob <double>
           Probability  of  generating  a  read  from the forward strand of a transcript. Set to 1 for a strand-
           specific protocol where all (upstream) reads are derived from the forward strand,  0  for  a  strand-
           specific  protocol  where  all (upstream) read are derived from the reverse strand, or 0.5 for a non-
           strand-specific protocol. (Default: 0.5)

       --fragment-length-min <int>
           Minimum read/insert length allowed. This  is  also  the  value  for  the  Bowtie/Bowtie2  -I  option.
           (Default: 1)

       --fragment-length-max <int>
           Maximum  read/insert  length  allowed.  This  is  also  the  value for the Bowtie/Bowtie 2 -X option.
           (Default: 1000)

       --fragment-length-mean <double>
           (single-end data only) The mean of the fragment  length  distribution,  which  is  assumed  to  be  a
           Gaussian. (Default: -1, which disables use of the fragment length distribution)

       --fragment-length-sd <double>
           (single-end  data  only) The standard deviation of the fragment length distribution, which is assumed
           to be a Gaussian.  (Default: 0, which assumes that all fragments are of the same length, given by the
           rounded value of --fragment-length-mean)

       --estimate-rspd
           Set this option if you want to estimate the  read  start  position  distribution  (RSPD)  from  data.
           Otherwise, RSEM will use a uniform RSPD. (Default: off)

       --num-rspd-bins <int>
           Number  of  bins  in the RSPD. Only relevant when '--estimate-rspd' is specified.  Use of the default
           setting is recommended. (Default: 20)

       --gibbs-burnin <int>
           The number of burn-in rounds for RSEM's Gibbs sampler. Each round passes over  the  entire  data  set
           once.  If  RSEM can use multiple threads, multiple Gibbs samplers will start at the same time and all
           samplers share the same burn-in number. (Default: 200)

       --gibbs-number-of-samples <int>
           The total number of count vectors RSEM will collect from its Gibbs samplers. (Default: 1000)

       --gibbs-sampling-gap <int>
           The number of rounds between two succinct count vectors RSEM collects.  If  the  count  vector  after
           round N is collected, the count vector after round N + <int> will also be collected. (Default: 1)

       --ci-credibility-level <double>
           The credibility level for credibility intervals. (Default: 0.95)

       --ci-memory <int>
           Maximum  size  (in memory, MB) of the auxiliary buffer used for computing credibility intervals (CI).
           (Default: 1024)

       --ci-number-of-samples-per-count-vector <int>
           The number of read generating probability vectors sampled per sampled  count  vector.  The  crebility
           intervals  are  calculated by first sampling P(C | D) and then sampling P(Theta | C) for each sampled
           count vector. This option controls how many Theta vectors  are  sampled  per  sampled  count  vector.
           (Default: 50)

       --keep-intermediate-files
           Keep temporary files generated by RSEM.  RSEM creates a temporary directory, 'sample_name.temp', into
           which  it  puts  all intermediate output files. If this directory already exists, RSEM overwrites all
           files generated by previous RSEM runs inside of it. By default, after RSEM  finishes,  the  temporary
           directory is deleted.  Set this option to prevent the deletion of this directory and the intermediate
           files inside of it. (Default: off)

       --temporary-folder <string>
           Set  where to put the temporary files generated by RSEM. If the folder specified does not exist, RSEM
           will try to create it. (Default: sample_name.temp)

       --time
           Output time consumed by each step of RSEM to 'sample_name.time'. (Default: off)

DESCRIPTION

       In its default mode, this program aligns input reads against a reference transcriptome  with  Bowtie  and
       calculates  expression  values  using  the  alignments.   RSEM assumes the data are single-end reads with
       quality scores, unless the '--paired-end' or '--no-qualities' options are specified. Alternatively, users
       can  use  STAR  to  align  reads  using   the   '--star'   option.   RSEM   has   provided   options   in
       'rsem-prepare-reference'  to  prepare  STAR's  genome  indices.  Users  may use an alternative aligner by
       specifying '--alignments', and providing an alignment file in SAM/BAM/CRAM format. However, users  should
       make  sure  that  they  align against the indices generated by 'rsem-prepare-reference' and the alignment
       file satisfies the requirements mentioned in ARGUMENTS section.

       One  simple  way  to  make  the  alignment  file  satisfying  RSEM's   requirements   is   to   use   the
       'convert-sam-for-rsem'  script.  This  script accepts SAM/BAM/CRAM files as input and outputs a BAM file.
       For example, type the following command to convert a SAM file, 'input.sam', to a ready-for-use BAM  file,
       'input_for_rsem.bam':

         convert-sam-for-rsem input.sam input_for_rsem

       For details, please refer to 'convert-sam-for-rsem's documentation page.

NOTES

       1. Users must run 'rsem-prepare-reference' with the appropriate reference before using this program.

       2. For single-end data, it is strongly recommended that the user provide the fragment length distribution
       parameters   (--fragment-length-mean   and   --fragment-length-sd).    For  paired-end  data,  RSEM  will
       automatically learn a fragment length distribution from the data.

       3. Some aligner parameters have default values different from their original settings.

       4. With the '--calc-pme' option, posterior mean estimates will  be  calculated  in  addition  to  maximum
       likelihood estimates.

       5. With the '--calc-ci' option, 95% credibility intervals and posterior mean estimates will be calculated
       in addition to maximum likelihood estimates.

       6.  The  temporary  directory  and  all  intermediate  files  will  be  removed when RSEM finishes unless
       '--keep-intermediate-files' is specified.

OUTPUT

       sample_name.isoforms.results
           File containing isoform level expression estimates. The first line 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  [posterior_mean_count
           posterior_standard_deviation_of_count   pme_TPM   pme_FPKM   IsoPct_from_pme_TPM   TPM_ci_lower_bound
           TPM_ci_upper_bound  TPM_coefficient_of_quartile_variation   FPKM_ci_lower_bound   FPKM_ci_upper_bound
           FPKM_coefficient_of_quartile_variation]

           Fields  are  separated  by  the  tab  character.  Fields  within  "[]" are optional. They will not be
           presented if neither '--calc-pme' nor '--calc-ci' is set.

           'transcript_id' is the transcript name of this transcript. 'gene_id' is the gene  name  of  the  gene
           which  this  transcript  belongs  to (denote this gene as its parent gene). If no gene information is
           provided, 'gene_id' and 'transcript_id' are the same.

           'length' is this transcript's sequence length  (poly(A)  tail  is  not  counted).  'effective_length'
           counts  only  the  positions  that  can  generate  a  valid  fragment.  If  no poly(A) tail is added,
           'effective_length' is equal to transcript length - mean fragment length  +  1.  If  one  transcript's
           effective  length is less than 1, this transcript's both effective length and abundance estimates are
           set to 0.

           'expected_count' is the sum of the posterior probability of each read comes from this transcript over
           all reads. Because 1) each read aligning to this transcript has a probability of being generated from
           background noise; 2) RSEM may filter some alignable low quality reads, the sum of expected counts for
           all transcript are generally less than the total number of reads aligned.

           'TPM' stands for Transcripts Per Million. It is a relative measure of transcript abundance.  The  sum
           of  all  transcripts'  TPM  is  1 million. 'FPKM' stands for Fragments Per Kilobase of transcript per
           Million mapped reads. It is another relative measure of transcript abundance. If we define  l_bar  be
           the mean transcript length in a sample, which can be calculated as

           l_bar = \sum_i TPM_i / 10^6 * effective_length_i (i goes through every transcript),

           the following equation is hold:

           FPKM_i = 10^3 / l_bar * TPM_i.

           We can see that the sum of FPKM is not a constant across samples.

           'IsoPct'  stands for isoform percentage. It is the percentage of this transcript's abandunce over its
           parent gene's abandunce. If its parent gene has only one isoform  or  the  gene  information  is  not
           provided, this field will be set to 100.

           '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% credibility intervals for TPM and FPKM values. The bounds are
           inclusive (i.e. [l, u]).

           'TPM_coefficient_of_quartile_variation' and 'FPKM_coefficient_of_quartile_variation' are coefficients
           of quartile variation (CQV) for TPM and FPKM values. CQV is a  robust  way  of  measuring  the  ratio
           between the standard deviation and the mean. It is defined as

           CQV := (Q3 - Q1) / (Q3 + Q1),

           where Q1 and Q3 are the first and third quartiles.

       sample_name.genes.results
           File  containing  gene  level expression estimates. The first line 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  [posterior_mean_count
           posterior_standard_deviation_of_count    pme_TPM   pme_FPKM   TPM_ci_lower_bound   TPM_ci_upper_bound
           TPM_coefficient_of_quartile_variation             FPKM_ci_lower_bound             FPKM_ci_upper_bound
           FPKM_coefficient_of_quartile_variation]

           Fields  are  separated  by  the  tab  character.  Fields  within  "[]" are optional. They will not be
           presented if neither '--calc-pme' nor '--calc-ci' is set.

           'transcript_id(s)' is a comma-separated list of transcript_ids belonging to this  gene.  If  no  gene
           information is provided, 'gene_id' and 'transcript_id(s)' are identical (the 'transcript_id').

           A  gene's  'length'  and  'effective_length'  are defined as the weighted average of its transcripts'
           lengths and effective lengths (weighted by 'IsoPct'). A gene's abundance estimates are just  the  sum
           of its transcripts' abundance estimates.

       sample_name.alleles.results
           Only generated when the RSEM references are built with allele-specific transcripts.

           This  file contains allele level expression estimates for allele-specific expression calculation. The
           first line 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   [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
           TPM_coefficient_of_quartile_variation             FPKM_ci_lower_bound             FPKM_ci_upper_bound
           FPKM_coefficient_of_quartile_variation]

           Fields  are  separated  by  the  tab  character.  Fields  within  "[]" are optional. They will not be
           presented if neither '--calc-pme' nor '--calc-ci' is set.

           'allele_id' is the allele-specific name of this allele-specific transcript.

           'AlleleIsoPct' stands for allele-specific percentage on isoform level. It is the percentage  of  this
           allele-specific  transcript's  abundance  over  its  parent  transcript's  abundance.  If  its parent
           transcript has only one allele variant form, this field will be set to 100.

           'AlleleGenePct' stands for allele-specific percentage on gene level. It is  the  percentage  of  this
           allele-specific transcript's abundance over its parent gene's abundance.

           'AlleleIsoPct_from_pme_TPM'   and   'AlleleGenePct_from_pme_TPM'  have  similar  meanings.  They  are
           calculated based on posterior mean estimates.

           Please  note  that  if  this  file  is  present,  the  fields  'length'  and  'effective_length'   in
           'sample_name.isoforms.results'  should  be  interpreted similarly as the corresponding definitions in
           'sample_name.genes.results'.

       sample_name.transcript.bam
           Only generated when --no-bam-output is not specified.

           'sample_name.transcript.bam' is a BAM-formatted file of read alignments  in  transcript  coordinates.
           The  MAPQ  field  of each alignment is set to min(100, floor(-10 * log10(1.0 - w) + 0.5)), where w is
           the posterior probability of that alignment being the true mapping of a read.  In addition, RSEM pads
           a new tag ZW:f:value, where value is a single precision floating number  representing  the  posterior
           probability.  Because  this  file  contains  all alignment lines produced by bowtie or user-specified
           aligners, it can also be used as a replacement of the aligner generated BAM/SAM file.

       sample_name.transcript.sorted.bam and sample_name.transcript.sorted.bam.bai
           Only generated when --no-bam-output is not specified and --sort-bam-by-coordinate is specified.

           'sample_name.transcript.sorted.bam' and 'sample_name.transcript.sorted.bam.bai' are  the  sorted  BAM
           file and indices generated by samtools (included in RSEM package).

       sample_name.genome.bam
           Only generated when --no-bam-output is not specified and --output-genome-bam is specified.

           'sample_name.genome.bam'  is  a  BAM-formatted  file  of  read  alignments  in  genomic  coordinates.
           Alignments of reads that have identical genomic coordinates (i.e., alignments to  different  isoforms
           that  share  the  same  genomic  region)  are  collapsed  into one alignment.  The MAPQ field of each
           alignment is set to min(100, floor(-10 * log10(1.0 - w) + 0.5)), where w is the posterior probability
           of that alignment being the true mapping of a read.  In addition, RSEM pads  a  new  tag  ZW:f:value,
           where  value  is  a  single  precision  floating number representing the posterior probability. If an
           alignment is spliced, a XS:A:value tag is also added, where value is either '+' or '-' indicating the
           strand of the transcript it aligns to.

       sample_name.genome.sorted.bam and sample_name.genome.sorted.bam.bai
           Only  generated  when  --no-bam-output   is   not   specified,   and   --sort-bam-by-coordinate   and
           --output-genome-bam are specified.

           'sample_name.genome.sorted.bam'  and  'sample_name.genome.sorted.bam.bai' are the sorted BAM file and
           indices generated by samtools (included in RSEM package).

       sample_name.time
           Only generated when --time is specified.

           It contains  time  (in  seconds)  consumed  by  aligning  reads,  estimating  expression  levels  and
           calculating credibility intervals.

       sample_name.stat
           This  is  a  folder  instead  of  a file. All model related statistics are stored in this folder. Use
           'rsem-plot-model' can generate plots using this folder.

           'sample_name.stat/sample_name.cnt' contains alignment statistics. The format  and  meanings  of  each
           field are described in 'cnt_file_description.txt' under RSEM directory.

           'sample_name.stat/sample_name.model'  stores  RNA-Seq  model  parameters  learned  from the data. The
           format and meanings of each filed of this file are described  in  'model_file_description.txt'  under
           RSEM directory.

EXAMPLES

       Assume the path to the bowtie executables is in the user's PATH environment variable. Reference files are
       under '/ref' with name 'mouse_125'.

       1)  '/data/mmliver.fq',  single-end  reads  with  quality  scores.  Quality scores are encoded as for 'GA
       pipeline version >= 1.3'. We want to use 8 threads and generate a genome BAM file. In addition,  we  want
       to append gene/transcript names to the result files:

        rsem-calculate-expression --phred64-quals \
                                  -p 8 \
                                  --append-names \
                                  --output-genome-bam \
                                  /data/mmliver.fq \
                                  /ref/mouse_125 \
                                  mmliver_single_quals

       2)  '/data/mmliver_1.fq'  and  '/data/mmliver_2.fq', paired-end reads with quality scores. Quality scores
       are in SANGER format. We want to use 8 threads and do not generate a genome BAM file:

        rsem-calculate-expression -p 8 \
                                  --paired-end \
                                  /data/mmliver_1.fq \
                                  /data/mmliver_2.fq \
                                  /ref/mouse_125 \
                                  mmliver_paired_end_quals

       3) '/data/mmliver.fa', single-end reads without quality scores. We want to use 8 threads:

        rsem-calculate-expression -p 8 \
                                  --no-qualities \
                                  /data/mmliver.fa \
                                  /ref/mouse_125 \
                                  mmliver_single_without_quals

       4) Data are the same as 1). This time we assume the bowtie executables are under '/sw/bowtie'. We want to
       take a fragment length distribution into consideration. We set the fragment length mean to  150  and  the
       standard  deviation  to 35. In addition to a BAM file, we also want to generate credibility intervals. We
       allow RSEM to use 1GB of memory for CI calculation:

        rsem-calculate-expression --bowtie-path /sw/bowtie \
                                  --phred64-quals \
                                  --fragment-length-mean 150.0 \
                                  --fragment-length-sd 35.0 \
                                  -p 8 \
                                  --output-genome-bam \
                                  --calc-ci \
                                  --ci-memory 1024 \
                                  /data/mmliver.fq \
                                  /ref/mouse_125 \
                                  mmliver_single_quals

       5) '/data/mmliver_paired_end_quals.bam', BAM-formatted  alignments  for  paired-end  reads  with  quality
       scores. We want to use 8 threads:

        rsem-calculate-expression --paired-end \
                                  --alignments \
                                  -p 8 \
                                  /data/mmliver_paired_end_quals.bam \
                                  /ref/mouse_125 \
                                  mmliver_paired_end_quals

       6)  '/data/mmliver_1.fq.gz'  and  '/data/mmliver_2.fq.gz',  paired-end reads with quality scores and read
       files are compressed by gzip. We want to use  STAR  to  aligned  reads  and  assume  STAR  executable  is
       '/sw/STAR'. Suppose we want to use 8 threads and do not generate a genome BAM file:

        rsem-calculate-expression --paired-end \
                                  --star \
                                  --star-path /sw/STAR \
                                  --gzipped-read-file \
                                  -p 8 \
                                  /data/mmliver_1.fq.gz \
                                  /data/mmliver_2.fq.gz \
                                  /ref/mouse_125 \
                                  mmliver_paired_end_quals

perl v5.22.2                                       2016-06-04                       RSEM-CALCULATE-EXPRESSION(1)