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NanoSim-H
=========
.. image:: https://travis-ci.org/karel-brinda/NanoSim-H.svg?branch=master
:target: https://travis-ci.org/karel-brinda/NanoSim-H
.. image:: https://img.shields.io/badge/install%20with-bioconda-brightgreen.svg?style=flat-square
:target: https://anaconda.org/bioconda/nanosim-h
.. image:: https://badge.fury.io/py/NanoSim-H.svg
:target: https://badge.fury.io/py/NanoSim-H
About
-----
NanoSim-H is a simulator of Oxford Nanopore reads that captures the technology-specific features of ONT data,
and allows for adjustments upon improvement of Nanopore sequencing technology.
NanoSim-H has been derived from `NanoSim <https://github.com/bcgsc/NanoSim>`_,
a software package developed by Chen Yang at `Canada's Michael Smith Genome Sciences Centre <http://www.bcgsc.ca/>`_.
The fork was created from version 1.0.1 and the versions of NanoSim-H and NanoSim are kept synchronized.
NanoSim-H is implemented using Python uses R for model fitting.
In silico reads can be simulated from a given reference genome using ``nanosim-h``.
The NanoSim-H package is distributed with several precomputed error profiles, but
additional profiles can be computed using the ``nanosim-h-train``.
The main improvements compared to NanoSim are:
* Support for Python 3
* Support for `RNF <https://www.ncbi.nlm.nih.gov/pubmed/26353839>`_ read names
* Installation from `PyPI <https://pypi.python.org/pypi/NanoSim-H/>`_
* Error profiles distributed with the main package
* Automatic testing using `Travis <https://travis-ci.org/karel-brinda/NanoSim-H>`_
* Reproducible simulations (setting a seed for PRG)
* Improved interface with new parameters (e.g., for merging all contigs) and a progress bar
* Several minor bugs fixed
Quick example
-------------
Simulation of 100 reads from an *E.coli genome*.
.. code-block:: bash
pip install --upgrade nanosim-h
curl "https://www.ncbi.nlm.nih.gov/sviewer/viewer.fcgi?db=nuccore&dopt=fasta&val=545778205&sendto=on" | \
nanosim-h -n 100 -
Installation
------------
**From** `BioConda <https://bioconda.github.io/>`_ **(recommended):**
.. code-block:: bash
conda install -y -c bioconda nanosim-h
**From** `PyPI <https://pypi.python.org/pypi>`_ **:**
.. code-block:: bash
pip install --upgrade nanosim-h
**From Github:**
.. code-block:: bash
git clone https://github.com/karel-brinda/nanosim-h
cd nanosim-h
pip install --upgrade .
or
.. code-block:: bash
git clone https://github.com/karel-brinda/nanosim-h
cd nanosim-h
python setup.py install
**Dependencies:**
For read simulation:
* `Python <http://python.org>`_ (2.7, 3.2 - 3.6)
* `Numpy <http://www.numpy.org/>`_
For computing new error profiles:
* `LAST <http://last.cbrc.jp/>`_ (tested with version 847)
* `R <https://www.r-project.org/>`_
When installed using Bioconda, all NanoSim-H dependencies get installed automatically.
When installed using PIP, all dependencies for read simulation are installed automatically.
Read simulation
---------------
Simulation stage takes a reference genome and possibly a read profile as input, and outputs simulated reads in FASTA format.
.. command: nanosim-h --help
.. code-block::
$ nanosim-h --help
usage: nanosim-h [-h] [-p str] [-o str] [-n int] [-u float] [-m float]
[-i float] [-d float] [-s int] [--circular] [--perfect]
[--merge-contigs] [--rnf] [--rnf-add-cigar] [--max-len int]
[--min-len int] [--kmer-bias int]
<reference.fa>
Program: NanoSim-H - a simulator of Oxford Nanopore reads.
Version: 1.1.0.0
Authors: Chen Yang <cheny@bcgsc.ca> - author of the original software package (NanoSim)
Karel Brinda <kbrinda@hsph.harvard.edu> - author of the NanoSim-H fork
positional arguments:
<reference.fa> reference genome (- for standard input)
optional arguments:
-h, --help show this help message and exit
-p str, --profile str
error profile - one of precomputed profiles
('ecoli_R7', 'ecoli_R7.3', 'ecoli_R9_1D',
'ecoli_R9_2D', 'ecoli_UCSC1b', 'yeast') or own
directory with an error profile [ecoli_R9_2D]
-o str, --out-pref str
prefix of output file [simulated]
-n int, --number int number of generated reads [10000]
-u float, --unalign-rate float
rate of unaligned reads [detect from the error
profile]
-m float, --mis-rate float
mismatch rate (weight tuning) [1.0]
-i float, --ins-rate float
insertion rate (weight tuning) [1.0]
-d float, --del-rate float
deletion rate (weight tuning) [1.0]
-s int, --seed int initial seed for the pseudorandom number generator (0
for random) [42]
--circular circular simulation (linear otherwise)
--perfect output perfect reads, no mutations
--merge-contigs merge contigs from the reference
--rnf use RNF format for read names
--rnf-add-cigar add cigar to RNF names (not fully debugged, yet)
--max-len int maximum read length [inf]
--min-len int minimum read length [50]
--kmer-bias int prohibits homopolymers with length >= n bases in
output reads [6]
Examples: nanosim-h --circular ecoli_ref.fasta
nanosim-h --circular --perfect ecoli_ref.fasta
nanosim-h -p yeast --kmer-bias 0 yeast_ref.fasta
Notice: the use of `max-len` and `min-len` will affect the read length distributions. If
the range between `max-len` and `min-len` is too small, the program will run slowlier accordingly.
.. end
**Examples:**
1. If you want to simulate reads from *E. coli* genome, then circular mode should be used because it is a circular genome.
``nanosim-h --circular Ecoli_ref.fasta``
2. If you want to simulate only perfect reads, i.e. no SNPs, or indels, just simulate the read length distribution.
``nanosimh-h --circular --perfect Ecoli_ref.fasta``
3. If you want to simulate reads from a *S. cerevisiae* genome with no *k*-mer bias, then linear mode should be chosen because it is a linear genome.
``nanosimh-h -p yeast --kmer-bias 0 yeast_ref.fasta``
**Output files:**
1. ``simulated.log`` – Log file for simulation process.
2. ``simulated_reads.fasta`` – FASTA file of simulated reads. Reads can contain information about how they were created either in RNF, or in the original NanoSim naming convention.
**RNF naming convention**
See the associated `RNF paper <https://www.ncbi.nlm.nih.gov/pubmed/26353839/>`_ and `RNF specification <karel-brinda.github.io/rnf-spec/>`_.
**NanoSim naming convention**
Each reads has "unaligned", "aligned", or "perfect" in the header determining their error rate. "unaligned" means that the reads have an error rate over 90% and cannot be aligned. "aligned" reads have the same error rate as training reads. "perfect" reads have no errors.
To explain the information in the header, we have two examples:
* ``>ref|NC-001137|-[chromosome=V]_468529_unaligned_0_F_0_3236_0``
All information before the first ``_`` are chromosome information. ``468529`` is the start position and *unaligned* suggesting it should be unaligned to the reference. The first ``0`` is the sequence index. ``F`` represents a forward strand. ``0_3236_0`` means that sequence length extracted from the reference is 3236 bases.
* ``>ref|NC-001143|-[chromosome=XI]_115406_aligned_16565_R_92_12710_2``
This is an aligned read coming from chromosome XI at position 115406. ``16565`` is the sequence index. `R` represents a reverse complement strand. ``92_12710_2`` means that this read has 92-base head region (cannot be aligned), followed by 12710 bases of middle region, and then 2-base tail region.
The information in the header can help users to locate the read easily.
3. ``simulated_errors`` – List of introduced errors.
The output contains error type, position, original bases and current bases.
Error profiles
--------------
Characterization stage takes a reference and a training read set in FASTA format as input. User can also provide their own alignment file in MAF format.
**Profiles distributed with NanoSim-H:**
* ``ecoli_R7``
* ``ecoli_R7.3``
* ``ecoli_R9_1D``
* ``ecoli_R9_2D`` (default error profile for read simulation)
* ``ecoli_UCSC1b``
* ``yeast``
**New error profiles:**
A new error profile can be obtained using the ``nanosim-h-train`` command.
.. command: nanosim-h-train --help
.. code-block::
$ nanosim-h-train --help
usage: nanosim-h-train [-h] [-i str] [-m str] [-b int] [--no-model-fit]
<reference.fa> <profile.dir>
Program: NanoSim-H-Train - compute an error profile for NanoSim-H.
Version: 1.1.0.0
Authors: Chen Yang <cheny@bcgsc.ca> - author of the original software package (NanoSim)
Karel Brinda <kbrinda@hsph.harvard.edu> - author of the NanoSim-H fork
positional arguments:
<reference.fa> reference genome of the training reads
<profile.dir> error profile dir
optional arguments:
-h, --help show this help message and exit
-i str, --infile str training ONT real reads, must be fasta files
-m str, --maf str user can provide their own alignment file, with maf
extension
-b int, --num-bins int
number of bins (for development) [20]
--no-model-fit no model fitting
.. end
**Files associated with an error profile:**
1. ``aligned_length_ecdf`` – Length distribution of aligned regions on aligned reads.
2. ``aligned_reads_ecdf`` – Length distribution of aligned reads.
3. ``align_ratio`` – Empirical distribution of align ratio of each read.
4. ``besthit.maf`` – The best alignment of each read based on length.
5. ``match.hist``, ``mis.hist``, ``ins.hist``, ``del.hist`` – Histograms of matches, mismatches, insertions, and deletions.
6. ``first_match.hist`` – Histogram of the first match length of each alignment.
7. ``error_markov_model`` – Markov model of error types.
8. ``ht_ratio`` – Empirical distribution of the head region vs total unaligned region.
9. ``training.maf`` – The output of LAST, alignment file in MAF format.
10. ``match_markov_model`` – Markov model of the length of matches (stretches of correct base calls).
11. ``model_profile`` – Fitted model for errors.
12. ``processed.maf`` – A re-formatted MAF file for user-provided alignment file.
13. ``unaligned_length_ecdf`` – Length distribution of unaligned reads
=========
.. image:: https://travis-ci.org/karel-brinda/NanoSim-H.svg?branch=master
:target: https://travis-ci.org/karel-brinda/NanoSim-H
.. image:: https://img.shields.io/badge/install%20with-bioconda-brightgreen.svg?style=flat-square
:target: https://anaconda.org/bioconda/nanosim-h
.. image:: https://badge.fury.io/py/NanoSim-H.svg
:target: https://badge.fury.io/py/NanoSim-H
About
-----
NanoSim-H is a simulator of Oxford Nanopore reads that captures the technology-specific features of ONT data,
and allows for adjustments upon improvement of Nanopore sequencing technology.
NanoSim-H has been derived from `NanoSim <https://github.com/bcgsc/NanoSim>`_,
a software package developed by Chen Yang at `Canada's Michael Smith Genome Sciences Centre <http://www.bcgsc.ca/>`_.
The fork was created from version 1.0.1 and the versions of NanoSim-H and NanoSim are kept synchronized.
NanoSim-H is implemented using Python uses R for model fitting.
In silico reads can be simulated from a given reference genome using ``nanosim-h``.
The NanoSim-H package is distributed with several precomputed error profiles, but
additional profiles can be computed using the ``nanosim-h-train``.
The main improvements compared to NanoSim are:
* Support for Python 3
* Support for `RNF <https://www.ncbi.nlm.nih.gov/pubmed/26353839>`_ read names
* Installation from `PyPI <https://pypi.python.org/pypi/NanoSim-H/>`_
* Error profiles distributed with the main package
* Automatic testing using `Travis <https://travis-ci.org/karel-brinda/NanoSim-H>`_
* Reproducible simulations (setting a seed for PRG)
* Improved interface with new parameters (e.g., for merging all contigs) and a progress bar
* Several minor bugs fixed
Quick example
-------------
Simulation of 100 reads from an *E.coli genome*.
.. code-block:: bash
pip install --upgrade nanosim-h
curl "https://www.ncbi.nlm.nih.gov/sviewer/viewer.fcgi?db=nuccore&dopt=fasta&val=545778205&sendto=on" | \
nanosim-h -n 100 -
Installation
------------
**From** `BioConda <https://bioconda.github.io/>`_ **(recommended):**
.. code-block:: bash
conda install -y -c bioconda nanosim-h
**From** `PyPI <https://pypi.python.org/pypi>`_ **:**
.. code-block:: bash
pip install --upgrade nanosim-h
**From Github:**
.. code-block:: bash
git clone https://github.com/karel-brinda/nanosim-h
cd nanosim-h
pip install --upgrade .
or
.. code-block:: bash
git clone https://github.com/karel-brinda/nanosim-h
cd nanosim-h
python setup.py install
**Dependencies:**
For read simulation:
* `Python <http://python.org>`_ (2.7, 3.2 - 3.6)
* `Numpy <http://www.numpy.org/>`_
For computing new error profiles:
* `LAST <http://last.cbrc.jp/>`_ (tested with version 847)
* `R <https://www.r-project.org/>`_
When installed using Bioconda, all NanoSim-H dependencies get installed automatically.
When installed using PIP, all dependencies for read simulation are installed automatically.
Read simulation
---------------
Simulation stage takes a reference genome and possibly a read profile as input, and outputs simulated reads in FASTA format.
.. command: nanosim-h --help
.. code-block::
$ nanosim-h --help
usage: nanosim-h [-h] [-p str] [-o str] [-n int] [-u float] [-m float]
[-i float] [-d float] [-s int] [--circular] [--perfect]
[--merge-contigs] [--rnf] [--rnf-add-cigar] [--max-len int]
[--min-len int] [--kmer-bias int]
<reference.fa>
Program: NanoSim-H - a simulator of Oxford Nanopore reads.
Version: 1.1.0.0
Authors: Chen Yang <cheny@bcgsc.ca> - author of the original software package (NanoSim)
Karel Brinda <kbrinda@hsph.harvard.edu> - author of the NanoSim-H fork
positional arguments:
<reference.fa> reference genome (- for standard input)
optional arguments:
-h, --help show this help message and exit
-p str, --profile str
error profile - one of precomputed profiles
('ecoli_R7', 'ecoli_R7.3', 'ecoli_R9_1D',
'ecoli_R9_2D', 'ecoli_UCSC1b', 'yeast') or own
directory with an error profile [ecoli_R9_2D]
-o str, --out-pref str
prefix of output file [simulated]
-n int, --number int number of generated reads [10000]
-u float, --unalign-rate float
rate of unaligned reads [detect from the error
profile]
-m float, --mis-rate float
mismatch rate (weight tuning) [1.0]
-i float, --ins-rate float
insertion rate (weight tuning) [1.0]
-d float, --del-rate float
deletion rate (weight tuning) [1.0]
-s int, --seed int initial seed for the pseudorandom number generator (0
for random) [42]
--circular circular simulation (linear otherwise)
--perfect output perfect reads, no mutations
--merge-contigs merge contigs from the reference
--rnf use RNF format for read names
--rnf-add-cigar add cigar to RNF names (not fully debugged, yet)
--max-len int maximum read length [inf]
--min-len int minimum read length [50]
--kmer-bias int prohibits homopolymers with length >= n bases in
output reads [6]
Examples: nanosim-h --circular ecoli_ref.fasta
nanosim-h --circular --perfect ecoli_ref.fasta
nanosim-h -p yeast --kmer-bias 0 yeast_ref.fasta
Notice: the use of `max-len` and `min-len` will affect the read length distributions. If
the range between `max-len` and `min-len` is too small, the program will run slowlier accordingly.
.. end
**Examples:**
1. If you want to simulate reads from *E. coli* genome, then circular mode should be used because it is a circular genome.
``nanosim-h --circular Ecoli_ref.fasta``
2. If you want to simulate only perfect reads, i.e. no SNPs, or indels, just simulate the read length distribution.
``nanosimh-h --circular --perfect Ecoli_ref.fasta``
3. If you want to simulate reads from a *S. cerevisiae* genome with no *k*-mer bias, then linear mode should be chosen because it is a linear genome.
``nanosimh-h -p yeast --kmer-bias 0 yeast_ref.fasta``
**Output files:**
1. ``simulated.log`` – Log file for simulation process.
2. ``simulated_reads.fasta`` – FASTA file of simulated reads. Reads can contain information about how they were created either in RNF, or in the original NanoSim naming convention.
**RNF naming convention**
See the associated `RNF paper <https://www.ncbi.nlm.nih.gov/pubmed/26353839/>`_ and `RNF specification <karel-brinda.github.io/rnf-spec/>`_.
**NanoSim naming convention**
Each reads has "unaligned", "aligned", or "perfect" in the header determining their error rate. "unaligned" means that the reads have an error rate over 90% and cannot be aligned. "aligned" reads have the same error rate as training reads. "perfect" reads have no errors.
To explain the information in the header, we have two examples:
* ``>ref|NC-001137|-[chromosome=V]_468529_unaligned_0_F_0_3236_0``
All information before the first ``_`` are chromosome information. ``468529`` is the start position and *unaligned* suggesting it should be unaligned to the reference. The first ``0`` is the sequence index. ``F`` represents a forward strand. ``0_3236_0`` means that sequence length extracted from the reference is 3236 bases.
* ``>ref|NC-001143|-[chromosome=XI]_115406_aligned_16565_R_92_12710_2``
This is an aligned read coming from chromosome XI at position 115406. ``16565`` is the sequence index. `R` represents a reverse complement strand. ``92_12710_2`` means that this read has 92-base head region (cannot be aligned), followed by 12710 bases of middle region, and then 2-base tail region.
The information in the header can help users to locate the read easily.
3. ``simulated_errors`` – List of introduced errors.
The output contains error type, position, original bases and current bases.
Error profiles
--------------
Characterization stage takes a reference and a training read set in FASTA format as input. User can also provide their own alignment file in MAF format.
**Profiles distributed with NanoSim-H:**
* ``ecoli_R7``
* ``ecoli_R7.3``
* ``ecoli_R9_1D``
* ``ecoli_R9_2D`` (default error profile for read simulation)
* ``ecoli_UCSC1b``
* ``yeast``
**New error profiles:**
A new error profile can be obtained using the ``nanosim-h-train`` command.
.. command: nanosim-h-train --help
.. code-block::
$ nanosim-h-train --help
usage: nanosim-h-train [-h] [-i str] [-m str] [-b int] [--no-model-fit]
<reference.fa> <profile.dir>
Program: NanoSim-H-Train - compute an error profile for NanoSim-H.
Version: 1.1.0.0
Authors: Chen Yang <cheny@bcgsc.ca> - author of the original software package (NanoSim)
Karel Brinda <kbrinda@hsph.harvard.edu> - author of the NanoSim-H fork
positional arguments:
<reference.fa> reference genome of the training reads
<profile.dir> error profile dir
optional arguments:
-h, --help show this help message and exit
-i str, --infile str training ONT real reads, must be fasta files
-m str, --maf str user can provide their own alignment file, with maf
extension
-b int, --num-bins int
number of bins (for development) [20]
--no-model-fit no model fitting
.. end
**Files associated with an error profile:**
1. ``aligned_length_ecdf`` – Length distribution of aligned regions on aligned reads.
2. ``aligned_reads_ecdf`` – Length distribution of aligned reads.
3. ``align_ratio`` – Empirical distribution of align ratio of each read.
4. ``besthit.maf`` – The best alignment of each read based on length.
5. ``match.hist``, ``mis.hist``, ``ins.hist``, ``del.hist`` – Histograms of matches, mismatches, insertions, and deletions.
6. ``first_match.hist`` – Histogram of the first match length of each alignment.
7. ``error_markov_model`` – Markov model of error types.
8. ``ht_ratio`` – Empirical distribution of the head region vs total unaligned region.
9. ``training.maf`` – The output of LAST, alignment file in MAF format.
10. ``match_markov_model`` – Markov model of the length of matches (stretches of correct base calls).
11. ``model_profile`` – Fitted model for errors.
12. ``processed.maf`` – A re-formatted MAF file for user-provided alignment file.
13. ``unaligned_length_ecdf`` – Length distribution of unaligned reads
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