# Installation¶

The SNP Pipeline software package consists of python scripts and shell scripts with dependencies on executable programs launched by the scripts.

## Step 1 - Operating System Requirements¶

The SNP Pipeline runs in a Linux environment. It has been tested on the following platforms:

• Red Hat
• CentOS
• Ubuntu

## Step 2 - Executable Software Dependencies¶

You should have the following software installed before using the SNP Pipeline.

Software Tested Version Description
Bowtie2 2.3.4.1 A tool for aligning reads to long reference sequences
SMALT 0.7.6 A tool for aligning reads to long reference sequences
SAMtools 1.8 Utilities for manipulating alignments in the SAM format
Picard 2.18.4 A set of tools for manipulating sequence data
GATK 3.8-1-0 Variant discovery and genotyping tools
VarScan 2.3.9 A tool to detect variants in NGS data
tabix 1.8 A generic indexer for tab-delimited genome position files
bgzip 1.8 Part of the tabix package, bgzip is a block compression utility
BcfTools 1.8 Utilities for variant calling and manipulating VCFs and BCFs
fastq-dump 2.8.1 An SRA Toolkit utility for fetching samples from NCBI SRA

Note: the versions above are tested and known to work together. Other versions may also work.

Note: you will need either Bowtie2 or SMALT. You do not have to install both. However, the included result files were generated with Bowtie2. Your results may differ when using SMALT.

Note: Picard is required when removing deplicate reads and when realigning reads around indels. Both of these functions are enabled by default, but can be disabled in the configuration file.

Note: GATK is required when realigning reads around indels, which is enabled by default, but can be disabled in the configuration file.

## Step 3 - Environment Variables¶

Define the CLASSPATH environment variable to specify the location of the Picard, VarScan, and GATK jar files. Add the following lines (or something similiar) to your .bashrc file:

export CLASSPATH=~/software/varscan.v2.3.9/VarScan.jar:$CLASSPATH export CLASSPATH=~/software/picard/picard.jar:$CLASSPATH
export CLASSPATH=~/software/GenomeAnalysisTK-3.8-1-0-gf15c1c3ef/GenomeAnalysisTK.jar:$CLASSPATH ## Step 4 - Python¶ The SNP pipeline is compatible with python version 2.6, 2.7, 3.3, 3.4, and 3.5. The pipeline has not been tested on other python versions. If you do not already have python installed, you should install version 2.7. You can either build from source or install a precompiled version with your Linux package manager. ## Step 5 - Pip¶ This can be a troublesome installation step – proceed with caution. The pip tool is used to install python packages including the snp-pipeline and other packages used by the snp-pipeline. Some newer versions of Python include pip. Check to see if pip is already installed:$ pip -V

If pip is not already installed, proceed as follows:

$python get-pip.py --user Note: avoid using sudo when installing pip. Some users have experienced problems installing and loading packages when pip is installed using sudo. ## Step 6 - Python Package Dependencies¶ For the most part, the installer automatically installs the necessary python packages used by snp-pipeline. However, not all python packages can be reliably installed automatically. The packages listed below may need to be manually installed if automatic installation fails. You can either install these packages now, or hope for the best and manually install later if the automatic installation fails. • Biopython, a set of tools for biological computation written in Python. ## Step 7 - Install the SNP Pipeline Python Package¶ There is more than one way to install the SNP Pipeline depending on whether you intend to work with the source code or just run it. ### Installation Method 1 for Most Users¶ This is the recommended installation method for new users. If you want to run the software without viewing or changing the source code, follow the instructions below. At the command line:$ pip install --user snp-pipeline

Update your .bashrc file with the path to user-installed python packages:

export PATH=~/.local/bin:$PATH Or, if you have virtualenvwrapper installed:$ mkvirtualenv snp-pipeline

## Uninstalling SNP Pipeline¶

If you installed with pip, you can uninstall from the command line:

$pip uninstall snp-pipeline ## Tips¶ There is a dependency on the python psutil package. Pip will attempt to install the psutil package automatically when installing snp-pipeline. If it fails with an error message about missing Python.h, you will need to manually install the python-dev package. In Ubuntu, use this command:$ sudo apt-get install python-dev

You may need to upgrade your Java Runtime Environment (JRE) to run Picard.