X-Git-Url: https://git.donarmstrong.com/?a=blobdiff_plain;f=README.md;h=f8c4e2709436f97e446ac3dbbda2306818df70f4;hb=90df7a1408511063de96e29658ffa289d43cc0bb;hp=0a76a6f06ab54afb865619ae30dafb464e2b1e89;hpb=3a69384beb61e14ce2830191538a6a26bb51d929;p=rsem.git
diff --git a/README.md b/README.md
index 0a76a6f..f8c4e27 100644
--- a/README.md
+++ b/README.md
@@ -18,7 +18,7 @@ Table of Contents
* * *
-
Introduction
+## Introduction
RSEM is a software package for estimating gene and isoform expression
levels from RNA-Seq data. The new RSEM package (rsem-1.x) provides an
@@ -27,9 +27,9 @@ the EM algorithm, single-end and paired-end read data, quality scores,
variable-length reads and RSPD estimation. It can also generate
genomic-coordinate BAM files and UCSC wiggle files for visualization. In
addition, it provides posterior mean and 95% credibility interval
-estimates for expression levels.
+estimates for expression levels.
-Compilation & Installation
+## Compilation & Installation
To compile RSEM, simply run
@@ -40,10 +40,14 @@ variable.
### Prerequisites
+C++ and Perl are required to be installed.
+
To take advantage of RSEM's built-in support for the Bowtie alignment
program, you must have [Bowtie](http://bowtie-bio.sourceforge.net) installed.
-Usage
+If you want to plot model learned by RSEM, you should also install R.
+
+## Usage
### I. Preparing Reference Sequences
@@ -62,7 +66,7 @@ To prepare the reference sequences, you should run the
rsem-prepare-reference --help
to get usage information or visit the [rsem-prepare-reference
-documentation page](rsem-prepare-reference.html).
+documentation page](http://deweylab.biostat.wisc.edu/rsem/rsem-prepare-reference.html).
### II. Calculating Expression Values
@@ -72,7 +76,7 @@ To calculate expression values, you should run the
rsem-calculate-expression --help
to get usage information or visit the [rsem-calculate-expression
-documentation page](rsem-calculate-expression.html).
+documentation page](http://deweylab.biostat.wisc.edu/rsem/rsem-calculate-expression.html).
#### Calculating expression values from single-end data
@@ -96,6 +100,12 @@ and provide the SAM or BAM file as an argument. When using an
alternative aligner, you may also want to provide the --no-bowtie option
to rsem-prepare-reference so that the Bowtie indices are not built.
+However, please note that RSEM does ** not ** support gapped
+alignments. So make sure that your aligner does not produce alignments
+with intersions/deletions. Also, please make sure that you use
+'reference_name.idx.fa' , which is generated by RSEM, to build your
+aligner's indices.
+
### III. Visualization
RSEM contains a version of samtools in the 'sam' subdirectory. When
@@ -123,7 +133,35 @@ wiggle_name: the name the user wants to use for this wiggle plot
Refer to the [UCSC custom track help page](http://genome.ucsc.edu/goldenPath/help/customTrack.html).
-Example
+#### c) Visualize the model learned by RSEM
+
+RSEM provides an R script, 'rsem-plot-model', for visulazing the model learned.
+
+Usage:
+
+ rsem-plot-model sample_name outF
+
+sample_name: the name of the sample analyzed
+outF: the file name for plots generated from the model. It is a pdf file
+
+The plots generated depends on read type and user configuration. It
+may include fragment length distribution, mate length distribution,
+read start position distribution (RSPD), quality score vs observed
+quality given a reference base, position vs percentage of sequencing
+error given a reference base and histogram of reads with different
+number of alignments.
+
+fragment length distribution and mate length distribution: x-axis is fragment/mate length, y axis is the probability of generating a fragment/mate with the associated length
+
+RSPD: Read Start Position Distribution. x-axis is bin number, y-axis is the probability of each bin. RSPD can be used as an indicator of 3' bias
+
+Quality score vs. observed quality given a reference base: x-axis is Phred quality scores associated with data, y-axis is the "observed quality", Phred quality scores learned by RSEM from the data. Q = -10log_10(P), where Q is Phred quality score and P is the probability of sequencing error for a particular base
+
+Position vs. percentage sequencing error given a reference base: x-axis is position and y-axis is percentage sequencing error
+
+Histogram of reads with different number of alignments: x-axis is the number of alignments a read has and y-axis is the number of such reads. The inf in x-axis means number of reads filtered due to too many alignments
+
+## Example
Suppose we download the mouse genome from UCSC Genome Browser. We will
use a reference_name of 'mm9'. We have a FASTQ-formatted file,
@@ -143,7 +181,7 @@ The commands for this scenario are as follows:
rsem-calculate-expression --bowtie-path /sw/bowtie --phred64-quals --fragment-length-mean 150.0 --fragment-length-sd 35.0 -p 8 --out-bam --calc-ci --memory-allocate 1024 /data/mmliver.fq /ref/mm9 mmliver_single_quals
rsem-bam2wig mmliver_single_quals.sorted.bam mmliver_single_quals.sorted.wig mmliver_single_quals
-Simulation
+## Simulation
### Usage:
@@ -168,13 +206,11 @@ output_name_1.fq & output_name_2.fq if paired-end with quality score.
output_name.sim.isoforms.results, output_name.sim.genes.results : Results estimated based on sample values.
-Acknowledgements
+## Acknowledgements
-RSEM uses randomc.h and mersenne.cpp from
- for random number generation. RSEM
-also uses the [Boost C++](http://www.boost.org) and
+RSEM uses the [Boost C++](http://www.boost.org) and
[samtools](http://samtools.sourceforge.net) libraries.
-License
+## License
RSEM is licensed under the [GNU General Public License v3](http://www.gnu.org/licenses/gpl-3.0.html).