Annotation with Prokka

Prokka is a tool that facilitates the fast annotation of prokaryotic genomes.

The goals of this tutorial are to:

  • Install Prokka
  • Use Prokka to annotate our genomes

Installing Prokka

Download and extract the latest version of prokka:

cd ~/
tar -xvzf prokka-1.11.tar.gz

We also will need some dependencies such as bioperl:

sudo apt-get -y install bioperl libdatetime-perl libxml-simple-perl libdigest-md5-perl

and we need an XML package from perl

sudo bash
export PERL_EXTUTILS_AUTOINSTALL="--defaultdeps"
perl -MCPAN -e 'install "XML::Simple"'

Now, you should be able to add Prokka to your $PATH and set up the index for the sequence database:

export PATH=$PATH:$HOME/prokka-1.11/bin
prokka --setupdb

Prokka should be good to go now– you can check to make sure that all is well by typing prokka. This should print the help screen with all available options. You can find out more about Prokka databases here.

Running Prokka

Make a new directory for the annotation:

cd ~/
mkdir annotation
cd annotation

Link the metagenome assembly file into this directory:

ln -fs ~/mapping/subset_assembly.fa .

Now it is time to run Prokka! There are tons of different ways to specialize the running of Prokka. We are going to keep it simple for now, though. It will take a little bit to run.

prokka subset_assembly.fa --outdir prokka_annotation --prefix metagG --metagenome

This will generate a new folder called prokka_annotation in which will be a series of files, which are detailed here.

In particular, we will be using the *.ffn file to assess the relative read coverage within our metagenomes across the predicted genomic regions.


  • What can I annotate with prokka?
  • Alternatives?
  • How do I submit my annotated files to Genbank? EBI??
  • Why is it called Prokka?

LICENSE: This documentation and all textual/graphic site content is released under Creative Commons - 0 (CC0) -- fork @ github.