Checking required software

Before we begin, we will quickly go through the required software and datasets for this workshop. For those who are already command-line-skilled there will also be a possibility to install the Metaxa2 tool for rRNA finding, but this is not required to complete the workshop.

Programs used in this workshop

The following programs are used in this workshop:

Since we are going to use the plotting functionality of R, we need to login to Uppmax with X11 forwarding turned on. In the Unix/Linux terminal this is easily achieved by adding the -X (captal X) option. All programs but Metaxa2 are already installed, all you have to do is load the virtual environment for this workshop. Once you are logged in to the server run:

source /proj/g2014113/metagenomics/virt-env/mg-workshop/bin/activate

You deactivate the virtual environment with:

deactivate

NOTE: This is a python virtual environment. The binary folder of the virtual environment has symbolic links to all programs used in this workshop so you should be able to run those without problems.

Check all programs in one go with which

To check whether you have all programs installed in one go, you can use which to test for the following programs:

hmmsearch
transeq
R
blastall

Data and databases used in this workshop

In this workshop, we are (due to time constraints) going to use a simplified version of the Pfam database, including only protein families related to plasmid replication and maintenance. This database is pre-compiled and can be downloaded from http://microbiology.se/teach/scilife2014/pfam.tar.gz Download it using the following commands:

mkdir -p ~/Pfam
cd ~/Pfam
wget http://microbiology.se/teach/scilife2014/pfam.tar.gz
tar -xzvf pfam.tar.gz
cd ~

In addition, you will need to obtain the following data sets for the workshop:

/proj/g2014113/metagenomics/annotation/baltic1.fna
/proj/g2014113/metagenomics/annotation/baltic2.fna
/proj/g2014113/metagenomics/annotation/indian_lake.fna
/proj/g2014113/metagenomics/annotation/swedish_lake.fna

We are going to use two data sets from the Baltic Sea, one from a Swedish lake and one from an Indian lake contaminated with wastewater from pharmaceutical production. For the same of time, I have reduced the data sets in size dramatically prior to this workshop. You can create links to the above files using the ln -s <path> command. Use it on all the four data sets.

(Optional excercise) Install Metaxa2 by yourself

Follow these steps only if you want to install Metaxa2 by yourself. The code for Metaxa2 is available from http://microbiology.se/sw/Metaxa2_2.0rc3.tar.gz You can install Metaxa2 as follows:

# Create a src and a bin directory
mkdir -p ~/src
mkdir -p ~/bin

# Go to the source directory and download the Metaxa2 tarball
cd ~/src
wget http://microbiology.se/sw/Metaxa2_2.0rc3.tar.gz
tar -xzvf Metaxa2_2.0rc3.tar.gz
cd Metaxa2_2.0rc3

# Run the installation script
./install_metaxa2

# Try to run Metaxa2 (this should bring up the main options for the software)
metaxa2 -h

If this did not work, you can try this manual approach:

cd ~/src/Metaxa2_2.0rc3
cp -r metaxa2* ~/bin/

# Then try to run Metaxa2 again
metaxa2 -h

If this brings up the help message, you are all set!