On a Linux machine, download a copy of Sunbeam from our GitHub repository, and install. We do not currently support non-Linux environments.
git clone -b stable https://github.com/sunbeam-labs/sunbeam sunbeam-stable cd sunbeam-stable ./install.sh tests/run_tests.bash -e sunbeam
This installs Sunbeam and all its dependencies, including the Conda environment manager, if required. It then runs some tests to make sure everything was installed correctly.
If you’ve never installed Conda before, you’ll need to add it to your shell’s
path. If you’re running Bash (the most common terminal shell), the following
command will add it to your path:
PATH=$PATH:$HOME/miniconda3/bin' > ~/.bashrc
If you see “Tests failed”, check out our troubleshooting section or file an issue on our GitHub page.
Let’s say your sequencing reads live in a folder called
/sequencing/project/reads, with one or two files per sample (for single- and
paired-end sequencing, respectively). These files must be in gzipped FASTQ
Let’s create a new Sunbeam project (we’ll call it
source activate sunbeam sunbeam init my_project --data_fp /sequencing/project/reads
Sunbeam will create a new folder called
my_project and put two files
sunbeam_config.ymlcontains all the configuration parameters for each step of the Sunbeam pipeline.
samples.csvis a comma-separated list of samples that Sunbeam found the given data folder, along with absolute paths to their FASTQ files.
Right now we have everything we need to do basic quality-control and contig assembly. However, let’s go ahead and set up contaminant filtering and some basic taxonomy databases to make things interesting.
Sunbeam can align your reads to an arbitrary number of contaminant sequences or host genomes and remove reads that map above a given threshold.
To use this, make a folder containing all the target sequences in FASTA
format. The filenames should end in “fasta” to be recognized by Sunbeam. In your
sunbeam_config.yml file, edit the
host_fp: line in the
section to point to this folder.
Sunbeam can use Kraken to assign putative taxonomic identities to your
reads. While creating a Kraken database is beyond the scope of this guide,
pre-built ones are available at the Kraken homepage. Download or build one, then add the
path to the database under
Sunbeam can automatically BLAST your contigs against any number of nucleotide or protein databases and summarize the top hits. Download or create your BLAST databases, then add the paths to your config file, following the instructions on here: blastdbs.
If you’d like to map the reads against a set of reference genomes of interest,
follow the same method as for the host/contaminant sequences above. Make a
folder containing FASTA files for each reference genome, then add the path to
that folder in
After you’ve finished editing your config file, you’re ready to run Sunbeam:
sunbeam run --configfile my_project/sunbeam_config.yml
By default, this will do a lot, including trimming and quality-controlling your
reads, removing contaminant, host, and low-complexity sequences, assigning
read-level taxonomy, assembling the reads in each sample into contigs, and then
BLASTing those contigs against your databases. Each of these steps can also be run independently by adding arguments after the
sunbeam run command. See Running for more info.