Clone this repository into a desired directory: git clone https://git.hhu.de/synmibi/reciprocal_blast_pipeline.git. Open a shell and cd into that directory, then type: docker-compose up. This can take several minutes to complete. After the reciprocal_blast_pipeline_web and reciprocal_blast_pipeline_db container have started open a new shell and cd into the project directory, then type: docker exec -it reciprocal_blast_pipeline_web /bin/bash. This will open a custom shell of the web container. Type: panoptes. This will start the panoptes server. In future versions panoptes will start with an entrypoint script. Now you can access the application by typing 127.0.0.1:8080/blast_project/ into your browser.

If you run into an error concerning the Django's database connection, try to CTRL-C the container process and restart both containers with docker-compose up.

In order to start developing and using this application you need to download the application from the git-repository. There are two possibilities for installing the application either you install packages with the package distribution platform anaconda and other executables manually or you can install the application with the container virtualization platform docker. Installation via docker involves execution of the docker-compose.yml file with the command docker-compose up. This will download and install the desired images and software requirements defined in the Dockerfile and requirements.txt file. Additionally, the application gets copied into the docker container. This may take some time. The resulting docker web image has a size of 3.02 Gigabyte. In order to execute snakemake you need to start the monitoring service panoptes in the docker container. Open a shell and submit following command: docker exec -it reciprocal_blast_pipeline_web /bin/bash. This will open a custom shell of the web container. Within the web container you need to submit the command panoptes. For installing the application within a conda environment you need the anaconda distribution platform and the biosnakedjango-{system}.yml file for your specific system. You can install the required packages in a virtual environment by starting a shell and submitting the command: conda env create -f biosnakedjango-{system}.yml In order to activate the environment use the command: conda activate biosnakedjango. If you install the application on a windows operating system you cannot use the E-Direct tool (further information at the end of this section). If you want to use the PostgreSQL database you need the additional python package psycopg2 and an active database server on your local machine, that runs on the port 5432. Otherwise, you can change the database to SQLite3 by editing the DATABASE section in the settings.py file, or by replacing everything in the DATABASE section with:

DATABASES = {
    'default': {
        'ENGINE': 'django.db.backends.sqlite3',
        'NAME': os.path.join(BASE_DIR, 'db.sqlite3'),
    }
}

Afterwards, change your active working directory to the reciprocal_blast project root folder and type following commands: python manage.py makemigration and python manage.py migrate. These commands will directly create the needed database tables. In the same working directory you can start the server with the command python manage.py runserver 8080. Snakemake is executed with the --wms-monitor http://127.0.0.1:5000 flag which tells snakemake to communicate with the monitoring service panoptes. In order to allow the correct snakemake execution and communication with panoptes, open a new shell in the project directory, activate the biosnakedjango environment and start the panoptes server by submitting the command panoptes. The last installation step for the conda setup involves downloading and installing the BLAST command line application and databases, as well as the E-Direct tool. For installation of those executables follow the instructions that are given in the BLAST and E-Direct user manuals. If you want to develop on windows, you cannot install the E-Direct tool. You need to out-comment the active part of the function write_taxid_file() in the blast_execution.py file and replace it with the currently out commented part.

The recommended installation procedure is the Docker setup. Especially windows users should install the application with docker, thus automated deletion of project files will not work correctly on windows platforms and installation of NCBI E-Direct tools is only possible on Mac or Linux operating systems. Load the project folder reciprocal_blast into your development IDE. One potential IDE that has plugins for snakemake support is PyCharm. Docker volume sharing between the host and container application directory enables development of this application inside the container. During container start the application gets started in development mode, thus Django uses an state reloader, which automatically reloads the development server whenever application files are changed. You do not need to stop the server and start it again if you make any code changes.

If you want to develop without Docker you should change the database settings. Changes are applied in the reciprocal_blast/reciprocal_blast/settings.py file. Replace the current postgres database settings with the settings for the SQLite database. Those settings are out commented in the relevant part of the settings file. If you have an active PostgreSQL server on your local machine you can use PostgreSQL. Additionally, various other databases are supported by Django.

Keep in mind that django is a MVT framework, which has its own markup language for designing templates and an own class based database monitoring. In addition to models and templates the core hook for execution of background tasks such as the execution of snakemake is the view.py file. Functions in this file will process the individual url specific HTTP GET and POST methods. This aspect is dealt with in more detail in the Django section of this readme and on the django project website).

HTML template files are rendered with Bootstrap 4, which is an HTML, CSS and JavaScript framework primarily for designing websites. A good overview of all design elements and options Bootstrap offers can be found on the Bootstrap project page.

Views are triggered in combination with the urls.py and views.py file. HTTP POST methods are connected to form validation. Form validation is defined in the forms.py file. Database transactions and file uploads are handled in the service.py file. Snakemake execution with previous configuration setup is handled in the blast_execution.py file. The project_creation view executes functions that are in the project_creation.py file which utilize functions of the service.py file.

Django is an open source web framework written in Python that follows a model-view-template (MVT framework) scheme, which allows a rapid and robust implementation of web applications. Django models allow an easy class based modeling of database tables. Model classes can be designed in the models.py file. Generally each model class maps to one appropriate database table. The underlying database is managed in the settings.py file. For this project a SQLite3 database for the local development and a PostgreSQL database for the development with docker containers is used. SQLite3 and PostgreSQL are relational database management systems that follow SQL (Structured-Query-Language) database engine specifications. The django models are directly linked to the data that is needed to execute the pipeline. In this work, the core model is the BlastProject class. Instances of this class are used as foreign keys for all other necessary models, e.g. ForwardBlastSettings, BackwardBlastSettings or Genomes, that are linked to the project. With an instance of the BlastProject class it is possible to get all relevant information for conducting the reciprocal BLAST analysis. The views manage the HTTP methods received by the server from user requests. Views are written in the views.py file, each view function has its own distinct responsibility such as the project_details view, which receives an user request and project_id and returns the current project-specific detail template. In turn, this detail template displays all of the project-specific data, such as the query sequence file and database names and the blast settings. Furthermore, if the pipeline execution finishes this view will present the resulting output data. Django will choose and execute a view function based on examining the URL that is requested, particularly on examining the part of the URL that comes after the domain name. Active URLs are stored within the urls.py files of your current project, hence all of the URLs can be found in the blast_project/urls.py file. A detailed overview is referenced in the user guidelines. Other project settings are managed within the reciprocal_blast/settings.py file.

The recommended installation procedure is the Docker setup. Especially windows users should install the application with docker, thus automated deletion of project files will not work correctly on windows platforms and installation of NCBI E-Direct tools is only possible on Mac or Linux operating systems. In order to enable the use of the preformatted nr database you have to download and decompress the database files from the NCBI FTP server . Additionally you have to download the taxonomy database (taxdb) in order to get taxonomic information in your blast output files. You can either download the files manually or use the update_blastdb.pl perl script that is provided by the BLAST executables. The size of the database is, after decompressing, roughly around 253 Gigabyte. Store the database in an appropriate directory and create a path variable called BLASTDB that points to the directory. If you choose the docker-compose.yaml file for the installation procedure you have to add two things to the container specifications: the database folder as a volume and an environment variable that points to this volume. This can be done in the docker-compose.yaml file with the volume and environment parameters. The application starts on port 8080, the postgres database starts with port 5432.

Once you have started the application you can visit the browser interface by typing and submitting localhost:8080/blast_project/ into your browser search bar. You need to register and create an account in order to use the application. After successful login you get redirected to the homepage of this application, which serves as a dashboard for your created projects. If you press the navigation bar button you can view a bunch of subsites, e.g. the project creation or the species taxonomic node checkup. Utilization of most subpages are self explaining.

During project creation you can either upload your own genome files or you can use the previously downloaded preformatted non-redundant database. All uploaded files should be in FASTA format. A more detailed user guideline is available on the github repository of this project.