This documentation is under active development, meaning that it can change over time as we refine it. Please email if you require assistance, or have suggestions to improve this documentation.


MASSIVE supports the bioinformatics, genomics and translational medicine community with storage and compute services. On these pages we will provide examples of workflows and software settings for running common bioinformatics software on M3.

Requesting an account on M3

If you are requesting an account on M3 and are working in collaboration with the Monash Bioinformatics Platform, please ensure you indicate this in the application and request that the appropriate Platform members are added to your M3 project. This will enable them to assist in your analysis.

Getting started

Importing the bioinformatics module environment

M3 has a number of bioinformatics packages available in the default set of modules. These include versions of bwa, bamtools, bcftools, bedtools, GATK, bcl2fastq, BEAST, BEAST2, bowtie, bowtie2, cufflinks, cytoscape, fastx-toolkit, kallisto, macs2, muscle, phyml, picard, qiime2, raxml, rstudio, samtools, star, sra-tools, subread, tophat, varscan, vep (this list shouldn’t be regarded as exhaustive !).

A software stack of additional packages (known as bio-ansible) is maintained by the Monash Bioinformatics Platform (MBP).

Tools are periodically added as required by the user community.

Modules maintained by MBP are installed at: /usr/local2/bioinformatics/

To access these additional tools, type:

source /usr/local2/bioinformatics/

This loads the bio-ansible modules into your environment alongside the default M3 modules. If you are using this frequently you might like to source this in your .profile / .bash_profile.

To list all modules:

module avail

You will see the additionals tools listed under the /usr/local2/bioinformatics/software/modules/bio section, followed by the default M3 modules.

Installing additional software with Bioconda

In addition to the pre-installed modules available on M3, Bioconda provides a streamlined way to install reproducible enviroments of bioinformatics tools in your home directory.

conda is already installed on M3 under the anaconda module.

module load anaconda

conda config --add channels r
conda config --add channels defaults
conda config --add channels conda-forge
conda config --add channels bioconda

These channels will now be listed in ~/.condarc and will be searched when installing packages by name.

Create a conda enviroment for your analysis / pipeline / project

Conda works by installing a set of pre-compiled tools and their dependencies in a self-contained ‘enviroments’ which you can switch between. Unlike modules loaded via module load, you can only have a single Conda environment active at one time.

To create an enviroment my_proj_env with a specific version of STAR, subread and asciigenome:

# Change this to your M3 project ID
export PROJECT=df22
export CONDA_ENVS=/projects/$PROJECT/$USER/conda_envs

mkdir -p $CONDA_ENVS

module load anaconda

conda create --yes -p $CONDA_ENVS/my_proj_env star=2.5.4a subread=1.5.2 asciigenome=1.12.0

# To use the environment
source activate $CONDA_ENVS/my_proj_env

# Leave the enviroment
source deactivate

You can search for packages on the commandline with: conda search <package_name>, or on the web using the Bioconda recipes list.

For further detail see the official Bioconda documentation.

Running the RNAsik RNA-seq pipeline on M3

The RNAsik pipeline runs on the BigDataScript workflow engine. A template BigDataScript config file pre-configured for M3 has been prepared, however you need to create a copy and modify the tmpDir setting to reflect your M3 project.

If the file ~/.bds/bds.config doesn’t exist, create a copy and edit the tmpDir setting like:

# Ensure the BigDataScript module is loaded
source /usr/local2/bioinformatics/
ml load BigDataScript

# Create a copy of the config file in your home directory
mkdir ~/.bds
cp $(which bds).config ~/.bds/

# Change this to your M3 project ID
export PROJECT=df22

# Edit the tmpDir setting to point to your scratch area
# (you can alternatively do this with vim / nano / emacs / $EDITOR)
sed -i 's/.*tmpDir.*/tmpDir = \/scratch\/${PROJECT}\/tmp\//' ~/.bds/bds.config

Create a SLURM sbatch script,, containing:

#SBATCH --account=${PROJECT}
#SBATCH --profile=All
#SBATCH --no-requeue
#SBATCH --ntasks=1
#SBATCH --ntasks-per-node=1
#SBATCH --cpus-per-task=2
#SBATCH --mem=2000
#SBATCH -t 3-0:00  # time limit (D-HH:MM)
#SBATCH --job-name=rnasik

# NOTE: The SBATCH options above only apply to the BigDataScript (bds) process.
# (This is why --mem and --cpus-per-task allocations are small since these only reflect the
# resources allocated to the BigDataScript pipeine runner, not STAR, picard etc).
# As part of running the pipeline, bds submits jobs to the queue with the appropriate (larger)
# --cpus-per-task and --mem flags. If you need to add SBATCH flags to every job submitted by
# bds (eg --account=myacct --reservation=special_nodes), add these flag to the
# clusterRunAdditionalArgs variable in the bds.config file.
# Default CPU and memory requirements are defined in bds.config and sik.config.

# Modify these variables as required

export REFERENCE_GENOMES="/scratch/${PROJECT}/references"
export FASTA_REF="${REFERENCE_GENOMES}/iGenomes/Mus_musculus/Ensembl/GRCm38/Sequence/WholeGenomeFasta/genome.fa"
export GTF_FILE="${REFERENCE_GENOMES}/iGenomes/Mus_musculus/Ensembl/GRCm38/Annotation/Genes/genes.gtf"
export FASTQ_DIR="./fastqs"
export SIK_VERSION="b55f2c7"

source "/usr/local2/bioinformatics/"

ml unload perl
ml load BigDataScript
ml load RNAsik-pipe/${SIK_VERSION}
ml load R
ml load python/3.6.2
ml load bwa
ml load hisat2
ml load bedtools2/2.25.0
ml load qualimap/v2.2.1
ml load gcc/6.1.0
ml load multiqc/1.4
ml load picard/2.18.0

ml list

# A precaution to ensure Java app wrapper scripts control -Xmx etc

# BigDataScript sometimes needs ParallelGCThreads set, but we don't want jobs
# to inherit these JVM settings.
# export _JAVA_OPTIONS=-Xms256M -Xmx728M -XX:ParallelGCThreads=1

export BDS_CONFIG="${HOME}/.bds/bds.config"

# /usr/local2/bioinformatics/software/apps/RNAsik-pipe-${SIK_VERSION}
export SIK_ORIGIN="$(dirname $(which RNAsik))/../"

# Run RNAsik bypassing the usual wrapper script so that we can specify additional options
bds -c ${BDS_CONFIG} -log -reportHtml "${SIK_ORIGIN}/src/RNAsik.bds" \
    -configFile "${SIK_ORIGIN}/configs/sik.config" \
    -fastaRef ${FASTA_REF} \
    -fqDir ${FASTQ_DIR} \
    -gtfFile ${GTF_FILE} \
    -align star \
    -counts \
    -all \
    -extn ".fastq.gz" \
    -paired \
    -pairIds "_R1,_R2"

Run like:



Q : You have version xx and I need version YY, how do I get the software?

A : You should consider installing the software in your home directory. The Bioconda project helps streamline this process with many pre-packaged tools for bioinformatics. If you are unable to install the version you need, please contact the helpdesk