#Website (http://sanger-pathogens.github.io/Roary)

#Roary the pan genome pipeline

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.

##Current citation "Roary: Rapid large-scale prokaryote pan genome analysis", Andrew J. Page, Carla A. Cummins, Martin Hunt, Vanessa K. Wong, Sandra Reuter, Matthew T. G. Holden, Maria Fookes, Jacqueline A. Keane, Julian Parkhill, bioRxiv doi: http://dx.doi.org/10.1101/019315

##Read the draft paper Roary: Rapid large-scale prokaryote pan genome analysis

##Input Roary takes annotated assemblies as input in GFF3 format, such as those produced by Prokka.

##Installation - Ubuntu/Debian Assuming you have root on your system, all the dependancies can be installed using apt and cpanm (only tested on Ubuntu 14.04).

sudo apt-get install bedtools cd-hit ncbi-blast+ mcl parallel cpanminus prank mafft exonerate
sudo cpanm -f Bio::Roary

###Older versions of Ubuntu/Debian (12.04 and below) Assuming you are running BASH, run this script, then copy and paste the last few lines into your BASH profile, as per the instructions. Not all the packages Roary requires are available on older versions of Ubuntu/Debian or the versions dont support features Roary requires. So this script will build them from source in the current working directory and install missing dependancies using apt and cpanm. This script is run automatically by our continous integration server which runs on Ubuntu 12.04.

./install_dependencies.sh

##Installation - OSX using homebrew and Linux using linuxbrew Assuming you have homebrew (OSX) or linuxbrew (Linux) setup and installed on your system:

brew tap homebrew/science
brew install bedtools cd-hit blast mcl parallel prank mafft exonerate
cpanm -f Bio::Roary

##Installation - With bundled binaries

###Download Download the latest software from https://github.com/sanger-pathogens/Roary/tarball/master

###Extract Choose somewhere to put it, for example in your home directory (no root access required):

cd $HOME
tar zxvf sanger-pathogens-Roary-xxxxxx.tar.gz
ls Roary-*

###Add to your Environment Add the following lines to your $HOME/.bashrc file, or to /etc/profile.d/roary.sh to make it available to all users:

export PATH=$PATH:$HOME/Roary-x.x.x/bin
export PERL5LIB=$PERL5LIB:$HOME/Roary-x.x.x/lib

###Install perl dependancies

cpanm Array::Utils BioPerl Exception::Class File::Find::Rule File::Grep File::Slurp::Tiny Graph Moose Moose::Role Text::CSV Log::Log4perl File::Which Graph::Writer::Dot Test::Files

##Installation - Ancient versions of Linux If none of the above options work, you'll have to install the depedancies from source or from your distributions packaging system. You should probably ask your system administrator for assistance if you havent done this kind of thing before.

##Installation - with Windows 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. You need to first install VirtualBox, then load the virtual machine, using the 'File -> Import Appliance' menu option. The root password is 'manager'.

ftp://ftp.sanger.ac.uk/pub/pathogens/pathogens-vm/pathogens-vm.latest.ova

#When things go wrong ###cdhit seg faults Old versions of cdhit have a bug, so you need to use at least version 4.6.1. The cdhit packages for Ubuntu 12.04 seem to be effected, so installing from the source is the only option.

###I installed the homebrew Kraken package and now theres an error when I run the tests or QC Theres a bug and you'll need to install it from source on older versions of OSX (like Mountain Lion).

###Why dont you bundle a Kraken database for the QC? Its massive (2.7GB) and changes as RefSeq is updated. The authors have prebuilt databases and details about how to make your own.