Align sequences and output the alignment in MSF format:

kalign -i BB11001.tfa -f msf  -o out.msf

Align sequences and output the alignment in clustal format:

kalign -i BB11001.tfa -f clu -o out.clu

Re-align sequences in an existing alignment:

kalign -i BB11001.msf  -o out.afa

Reformat existing alignment:

kalign -i BB11001.msf -r afa -o out.afa

Address of the bookmark: https://github.com/TimoLassmann/kalign

NucDiff provides information about the types of differences and their locations. It is possible to upload the results into genome browser for visualization and further inspection. It was written in Python and uses the NUCmer package from MUMmer[1] for sequence comparison.

Address of the bookmark: https://github.com/uio-cels/NucDiff

Key Features of BLAST:
1. Sequence Comparison: BLAST searches for local alignments between the query sequence and sequences in a database. It identifies regions of similarity, which can help infer functional and evolutionary relationships.

2. Speed and Efficiency: BLAST uses heuristic algorithms, making it faster than exhaustive search methods, suitable for large-scale database searches.

3. Versatility: There are several versions of BLAST for different types of sequence comparisons:
- blastn: Compares a nucleotide query sequence against a nucleotide sequence database.
- blastp: Compares a protein query sequence against a protein sequence database.
- blastx: Compares a nucleotide query sequence translated in all reading frames against a protein sequence database.
- tblastn: Compares a protein query sequence against a nucleotide sequence database translated in all reading frames.
- tblastx: Compares the six-frame translations of a nucleotide query sequence against the six-frame translations of a nucleotide sequence database.

4. Scoring and E-value: BLAST results are scored based on the quality and length of the alignments. The E-value (expect value) indicates the number of alignments one can expect to find by chance, with lower E-values representing more significant matches.

5. Output Formats: BLAST provides results in various formats, including plain text, HTML, XML, and JSON, making it adaptable for different types of analyses and integrations with other tools.

Applications of BLAST:
- Genomic Research: Identifying genes, understanding genetic diversity, and mapping genome sequences.
- Protein Function Prediction: Inferring the function of unknown proteins by comparing them to known protein sequences.
- Evolutionary Studies: Exploring evolutionary relationships between organisms by comparing their genetic material.
- Medical Research: Identifying pathogens, understanding disease mechanisms, and developing treatments by comparing sequences of interest.

Overall, BLAST is an essential tool in bioinformatics, offering a reliable and efficient way to analyze and interpret biological sequence data.

For further details about the underlying algorithm see the original publication:
MACSE: Multiple Alignment of Coding SEquences accounting for frameshifts and stop codons.
Vincent Ranwez, Sébastien Harispe, Frédéric Delsuc, Emmanuel JP Douzery
PLoS One 2011, 6(9): e22594.

Address of the bookmark: https://bioweb.supagro.inra.fr/macse/index.php?menu=releases

Prerequisites

• A fresh Vultr Cloud Compute instance with Ubuntu 18.04 and root access.

Step 1: Install Apache, MySQL, and PHP

Elgg requires MySQL, PHP, and a web server. Before you can install Elgg, you will need to install the Apache web server, MySQL, and PHP.

Update the repository list.

apt-get update

Install the Apache web server.

apt-get install apache2 -y

Install MySQL.

apt-get install mysql-server -y

Complete the MySQL installation by executing the following command.

/usr/bin/mysql_secure_installation

During the installation, you will be asked to enter a root password. Enter a secure password. This will be the MySQL root password.

Would you like to setup VALIDATE PASSWORD plugin? [Y/N] N
New password: password
Re-enter new password: password
Remove anonymous users? [Y/N] Y
Disallow root login remotely? [Y/N] Y
Remove test database and access to it? [Y/N] Y
Reload privilege tables now? [Y/N] Y

Install PHP 7.2, as well as the PHP modules required by Elgg.

apt-get install php7.2 libapache2-mod-php7.2 php7.2-common php7.2-sqlite3 php7.2-curl php7.2-intl php7.2-mbstring php7.2-xmlrpc php7.2-mysql php7.2-gd php7.2-xml php7.2-cli php7.2-zip -y

Step 2: Create a MySQL database for Elgg

Elgg will require a MySQL database. Log into the MySQL console.

mysql -u root -p

When prompted for a password, enter the MySQL root password you set in step 1. Once you are logged in to the MySQL console, create a new database.

CREATE DATABASE elgg;

Create a new MySQL user and grant it privileges to the newly created database. You can replace username and password with the username and password of your choice.

GRANT ALL PRIVILEGES on elgg.* to 'username'@'localhost' identified by 'password';
FLUSH PRIVILEGES;

Exit the MySQL console.

exit

Step 3: Download and Install Elgg

Download the latest version of Elgg.

cd /var/www/html
rm -r index.html
wget https://elgg.org/download/elgg-2.3.7.zip

Unzip the downloaded archive and move the files to the root of the Apache web server.

apt install unzip
unzip elgg-2.3.7.zip
mv ./elgg-2.3.7/* . && rm elgg-2.3.7.zip && rm -r elgg-2.3.7

Create a data directory for Elgg.

sudo mkdir -p /var/www/html/data

Set the appropriate file permissions.

sudo chown -R www-data:www-data /var/www/html/
sudo chmod -R 755 /var/www/html/

Step 4: Configure Apache for Elgg

Elgg requires the Apache rewrite module. Enable the Apache rewrite module.

sudo a2enmod rewrite

Create an Apache configuration file for the Elgg installation.

sudo nano /etc/apache2/sites-available/elgg.conf

Paste the following snippet to the file, replacing example.com with your own domain name.

<VirtualHost *:80>
     DocumentRoot /var/www/html/
     ServerName example.com
     <Directory /var/www/html/>
          Options FollowSymlinks
          AllowOverride All
          Require all granted
     </Directory>
     ErrorLog ${APACHE_LOG_DIR}/error.log
     CustomLog ${APACHE_LOG_DIR}/access.log combined
</VirtualHost>

Enable the configuration and restart the Apache server.

 sudo a2ensite elgg.conf
 sudo systemctl restart apache2.service

• nodes, which are labeled by sequences and ids
• edges, which connect two nodes via either of their respective ends
• paths, describe genomes, sequence alignments, and annotations (such as gene models and transcripts) as walks through nodes connected by edges

Address of the bookmark: https://github.com/vgteam/vg

Address of the bookmark: https://github.com/CSB5/INC-Seq

Address of the bookmark: https://github.com/bcgsc/ARCS/

more at https://ccb.jhu.edu/software/hisat2/index.shtml

Address of the bookmark: https://github.com/infphilo/hisat2