|
0
|
1 # Roary - The pan genome pipeline
|
|
|
2 Takes annotated assemblies in GFF3 format and calculates the pan genome.
|
|
|
3
|
|
|
4 PLEASE NOTE: we currently do not have the resources to provide support for Roary, so please do not expect a reply if you flag any issue.
|
|
|
5
|
|
|
6 [](http://unmaintained.tech/)
|
|
|
7 [](https://travis-ci.org/sanger-pathogens/Roary)
|
|
|
8 [](https://github.com/sanger-pathogens/roary/blob/master/GPL-LICENSE)
|
|
|
9 [](https://academic.oup.com/bioinformatics/article/31/22/3691/240757)
|
|
|
10 [](http://bioconda.github.io/recipes/roary/README.html)
|
|
|
11 [](https://quay.io/repository/biocontainers/roary)
|
|
|
12 [](https://hub.docker.com/r/sangerpathogens/roary)
|
|
|
13 [](https://hub.docker.com/r/sangerpathogens/roary)
|
|
|
14 [](https://codecov.io/gh/sanger-pathogens/roary)
|
|
|
15
|
|
|
16 ## Contents
|
|
|
17 * [Introduction](#introduction)
|
|
|
18 * [Installation](#installation)
|
|
|
19 * [Required dependencies](#required-dependencies)
|
|
|
20 * [Optional dependencies](#optional-dependencies)
|
|
|
21 * [Ubuntu/Debian](#ubuntudebian)
|
|
|
22 * [Debian Testing](#debian-testing)
|
|
|
23 * [Ubuntu 14\.04/16\.04](#ubuntu-14041604)
|
|
|
24 * [Ubuntu 12\.04](#ubuntu-1204)
|
|
|
25 * [Bioconda \- OSX/Linux](#bioconda---osxlinux)
|
|
|
26 * [Galaxy](#galaxy)
|
|
|
27 * [GNU Guix](#gnu-guix)
|
|
|
28 * [Virtual Machine \- OSX/Linux/Windows](#virtual-machine---osxlinuxwindows)
|
|
|
29 * [Docker \- OSX/Linux/Windows/Cloud](#docker---osxlinuxwindowscloud)
|
|
|
30 * [Installing from source (advanced Linux users only)](#installing-from-source-advanced-linux-users-only)
|
|
|
31 * [Ancient systems and versions of perl](#ancient-systems-and-versions-of-perl)
|
|
|
32 * [Running the tests](#running-the-tests)
|
|
|
33 * [Versions of software we test against](#versions-of-software-we-test-against)
|
|
|
34 * [Usage](#usage)
|
|
|
35 * [License](#license)
|
|
|
36 * [Feedback/Issues](#feedbackissues)
|
|
|
37 * [Citation](#citation)
|
|
|
38 * [Further Information](#further-information)
|
|
|
39
|
|
|
40 ## Introduction
|
|
|
41 Roary is a high speed stand alone pan genome pipeline, which takes annotated assemblies in GFF3 format (produced by Prokka) and calculates the pan genome. Using a standard desktop PC, it can analyse datasets with thousands of samples, something which is computationally infeasible with existing methods, without compromising the quality of the results. 128 samples can be analysed in under 1 hour using 1 GB of RAM and a single processor. To perform this analysis using existing methods would take weeks and hundreds of GB of RAM.
|
|
|
42
|
|
|
43 ## Installation
|
|
|
44 Roary has the following dependencies:
|
|
|
45
|
|
|
46 ### Required dependencies
|
|
|
47 * [bedtools](https://bedtools.readthedocs.io/en/latest/)
|
|
|
48 * [cd-hit](http://weizhongli-lab.org/cd-hit/)
|
|
|
49 * [ncbi-blast+](https://blast.ncbi.nlm.nih.gov/Blast.cgi?CMD=Web&PAGE_TYPE=BlastDocs&DOC_TYPE=Download)
|
|
|
50 * [mcl](https://micans.org/mcl/)
|
|
|
51 * [parallel](https://www.gnu.org/software/parallel/)
|
|
|
52 * [prank](http://wasabiapp.org/software/prank/)
|
|
|
53 * [mafft](https://mafft.cbrc.jp/alignment/software/)
|
|
|
54 * [fasttree](http://www.microbesonline.org/fasttree/)
|
|
|
55
|
|
|
56 ### Optional dependencies
|
|
|
57 * [kraken](http://ccb.jhu.edu/software/kraken/MANUAL.html)
|
|
|
58
|
|
|
59 There are a number of ways to install Roary and details are provided below. If you encounter an issue when installing Roary please contact your local system administrator.
|
|
|
60
|
|
|
61 ### Ubuntu/Debian
|
|
|
62 #### Debian Testing
|
|
|
63 ```
|
|
|
64 sudo apt-get install roary
|
|
|
65 ```
|
|
|
66
|
|
|
67 #### Ubuntu 14.04/16.04
|
|
|
68 All the dependancies can be installed using apt and cpanm. Root permissions are required. Ubuntu 16.04 contains a package for Roary but it is frozen at v3.6.0.
|
|
|
69
|
|
|
70 ```
|
|
|
71 sudo apt-get install bedtools cd-hit ncbi-blast+ mcl parallel cpanminus prank mafft fasttree
|
|
|
72 sudo cpanm -f Bio::Roary
|
|
|
73 ```
|
|
|
74
|
|
|
75 #### Ubuntu 12.04
|
|
|
76 Some of the software versions in apt are quite old so follow the instructions for Bioconda below.
|
|
|
77
|
|
|
78 ### Bioconda - OSX/Linux
|
|
|
79 Install conda. Then install bioconda and roary:
|
|
|
80
|
|
|
81 ```
|
|
|
82 conda config --add channels r
|
|
|
83 conda config --add channels defaults
|
|
|
84 conda config --add channels conda-forge
|
|
|
85 conda config --add channels bioconda
|
|
|
86 conda install roary
|
|
|
87 ```
|
|
|
88
|
|
|
89 ### Galaxy
|
|
|
90 Roary is available from the Galaxy toolshed (as is Prokka).
|
|
|
91
|
|
|
92 ### GNU Guix
|
|
|
93 Roary is included in [Guix](https://www.gnu.org/software/guix) and can be installed in the usual way:
|
|
|
94 ```
|
|
|
95 guix package --install roary
|
|
|
96 ```
|
|
|
97
|
|
|
98 ### Virtual Machine - OSX/Linux/Windows
|
|
|
99 Roary wont run natively on Windows but we have created virtual machine which has all of the software setup, including Prokka, along with the test datasets from the paper. It is based on [Bio-Linux 8](http://environmentalomics.org/bio-linux/). You need to first install [VirtualBox](https://www.virtualbox.org/), then load the virtual machine, using the 'File -> Import Appliance' menu option. The root password is 'manager'.
|
|
|
100
|
|
|
101 ftp://ftp.sanger.ac.uk/pub/pathogens/pathogens-vm/pathogens-vm.latest.ova
|
|
|
102
|
|
|
103 More importantly though, if you're trying to do bioinformatics on Windows, you're not going to get very far and you should seriously consider upgrading to Linux.
|
|
|
104
|
|
|
105 ### Docker - OSX/Linux/Windows/Cloud
|
|
|
106 We have a docker container which gets automatically built from the latest version of Roary in Debian Med. To install it:
|
|
|
107
|
|
|
108 ```
|
|
|
109 docker pull sangerpathogens/roary
|
|
|
110 ```
|
|
|
111
|
|
|
112 To use it you would use a command such as this (substituting in your directories), where your GFF files are assumed to be stored in /home/ubuntu/data:
|
|
|
113 ```
|
|
|
114 docker run --rm -it -v /home/ubuntu/data:/data sangerpathogens/roary roary -f /data /data/*.gff
|
|
|
115 ```
|
|
|
116
|
|
|
117 ### Installing from source (advanced Linux users only)
|
|
|
118 As a last resort you can install everything from source. This is for users with advanced Linux skills and we do not provide any support with this method since you have the skills to figure things out.
|
|
|
119 Download the latest software from (https://github.com/sanger-pathogens/Roary/tarball/master).
|
|
|
120
|
|
|
121 Choose somewhere to put it, for example in your home directory (no root access required):
|
|
|
122
|
|
|
123 ```
|
|
|
124 cd $HOME
|
|
|
125 tar zxvf sanger-pathogens-Roary-xxxxxx.tar.gz
|
|
|
126 ls Roary-*
|
|
|
127 ```
|
|
|
128
|
|
|
129 Add the following lines to your $HOME/.bashrc file, or to /etc/profile.d/roary.sh to make it available to all users:
|
|
|
130
|
|
|
131 ```
|
|
|
132 export PATH=$PATH:$HOME/Roary-x.x.x/bin
|
|
|
133 export PERL5LIB=$PERL5LIB:$HOME/Roary-x.x.x/lib
|
|
|
134 ```
|
|
|
135 Install the Perl dependencies:
|
|
|
136
|
|
|
137 ```
|
|
|
138 sudo cpanm Array::Utils Bio::Perl Exception::Class File::Basename File::Copy File::Find::Rule File::Grep File::Path File::Slurper File::Spec File::Temp File::Which FindBin Getopt::Long Graph Graph::Writer::Dot List::Util Log::Log4perl Moose Moose::Role Text::CSV PerlIO::utf8_strict Devel::OverloadInfo Digest::MD5::File
|
|
|
139 ```
|
|
|
140 Install the external dependances either from source or from your packaging system:
|
|
|
141 ```
|
|
|
142 bedtools cd-hit blast mcl GNUparallel prank mafft fasttree
|
|
|
143 ```
|
|
|
144
|
|
|
145 ### Ancient systems and versions of perl
|
|
|
146 The code will not work with perl 5.8 or below (pre-modern perl). We no longer test against 5.10 (released 2007) or 5.12 (released 2010). If you're running a very old verison of Linux, you're also in trouble.
|
|
|
147
|
|
|
148 ### Running the tests
|
|
|
149 The test can be run with dzil from the top level directory:
|
|
|
150
|
|
|
151 ```
|
|
|
152 dzil test
|
|
|
153 ```
|
|
|
154
|
|
|
155 ### Versions of software we test against
|
|
|
156 * Perl 5.14, 5.26
|
|
|
157 * cdhit 4.6.8
|
|
|
158 * ncbi blast+ 2.6.0
|
|
|
159 * mcl 14-137
|
|
|
160 * bedtools 2.27.1
|
|
|
161 * prank 140603
|
|
|
162 * GNU parallel 20170822, 20160722
|
|
|
163 * FastTree 2.1.9
|
|
|
164
|
|
|
165 ## Usage
|
|
|
166 ```
|
|
|
167 Usage: roary [options] *.gff
|
|
|
168
|
|
|
169 Options: -p INT number of threads [1]
|
|
|
170 -o STR clusters output filename [clustered_proteins]
|
|
|
171 -f STR output directory [.]
|
|
|
172 -e create a multiFASTA alignment of core genes using PRANK
|
|
|
173 -n fast core gene alignment with MAFFT, use with -e
|
|
|
174 -i minimum percentage identity for blastp [95]
|
|
|
175 -cd FLOAT percentage of isolates a gene must be in to be core [99]
|
|
|
176 -qc generate QC report with Kraken
|
|
|
177 -k STR path to Kraken database for QC, use with -qc
|
|
|
178 -a check dependancies and print versions
|
|
|
179 -b STR blastp executable [blastp]
|
|
|
180 -c STR mcl executable [mcl]
|
|
|
181 -d STR mcxdeblast executable [mcxdeblast]
|
|
|
182 -g INT maximum number of clusters [50000]
|
|
|
183 -m STR makeblastdb executable [makeblastdb]
|
|
|
184 -r create R plots, requires R and ggplot2
|
|
|
185 -s dont split paralogs
|
|
|
186 -t INT translation table [11]
|
|
|
187 -ap allow paralogs in core alignment
|
|
|
188 -z dont delete intermediate files
|
|
|
189 -v verbose output to STDOUT
|
|
|
190 -w print version and exit
|
|
|
191 -y add gene inference information to spreadsheet, doesnt work with -e
|
|
|
192 -iv STR Change the MCL inflation value [1.5]
|
|
|
193 -h this help message
|
|
|
194
|
|
|
195 Example: Quickly generate a core gene alignment using 8 threads
|
|
|
196 roary -e --mafft -p 8 *.gff
|
|
|
197
|
|
|
198 For further info see: http://sanger-pathogens.github.io/Roary/
|
|
|
199 ```
|
|
|
200 For further instructions on how to use the software, the input format and output formats, please see [the Roary website](http://sanger-pathogens.github.io/Roary).
|
|
|
201
|
|
|
202 ## License
|
|
|
203 Roary is free software, licensed under [GPLv3](https://github.com/sanger-pathogens/Roary/blob/master/GPL-LICENSE).
|
|
|
204
|
|
|
205 ## Feedback/Issues
|
|
|
206 We currently do not have the resources to provide support for Roary. However, the community might be able to help you out if you report any issues about usage of the software to the [issues page](https://github.com/sanger-pathogens/Roary/issues).
|
|
|
207
|
|
|
208 ## Citation
|
|
|
209 If you use this software please cite:
|
|
|
210
|
|
|
211 "Roary: Rapid large-scale prokaryote pan genome analysis",
|
|
|
212 Andrew J. Page, Carla A. Cummins, Martin Hunt, Vanessa K. Wong, Sandra Reuter, Matthew T. G. Holden, Maria Fookes, Daniel Falush, Jacqueline A. Keane, Julian Parkhill,
|
|
|
213 Bioinformatics, (2015). doi: http://dx.doi.org/10.1093/bioinformatics/btv421
|
|
|
214 [Roary: Rapid large-scale prokaryote pan genome analysis](http://dx.doi.org/10.1093/bioinformatics/btv421)
|
|
|
215
|
|
|
216 ## Further Information
|
|
|
217 For more information on this software see:
|
|
|
218 * [The Roary website](http://sanger-pathogens.github.io/Roary)
|
|
|
219 * [The Jupyter notebook tutorial](https://github.com/sanger-pathogens/pathogen-informatics-training)
|
