DIAMOND Tutorial


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Overview of DIAMOND

DIAMOND is a high-throughput bioinformatics software tool designed for aligning DNA reads or protein sequences against a protein reference database, such as the NCBI non-redundant (NR) database. It is renowned for its speed, being capable of performing alignments up to 20,000 times faster than the traditional BLASTX tool, while maintaining a high level of sensitivity.

The tool is particularly useful for analyzing Illumina reads ranging from 100 to 150 base pairs in length. In its fast mode, DIAMOND can process data approximately 20,000 times quicker than BLASTX, capturing about 80-90% of the matches that BLASTX would find with an e-value of at most 1e-5. When operating in sensitive mode, DIAMOND is about 2,500 times faster than BLASTX and finds more than 94% of all matches.

DIAMOND's impressive speed and sensitivity make it an invaluable resource for researchers working with large-scale genomic data, particularly in the fields of metagenomics and proteomics. It enables the rapid identification and analysis of protein-coding regions within vast amounts of sequence data, facilitating a deeper understanding of the functional implications of genetic information.

The tool was developed by Benjamin Buchfink, Chao Xie, and Daniel H. Huson, and its performance and capabilities were detailed in a publication in Nature Methods in 2015. Since its introduction, DIAMOND has become a staple in the bioinformatics community for protein alignment tasks.

For more detailed information and updates, users and developers are encouraged to refer to the original publication and the DIAMOND GitHub page, where the software can be downloaded and contributions to its development can be made.


To install DIAMOND, users should visit the DIAMOND GitHub page and follow the instructions provided. The installation process typically involves downloading the source code or binary files compatible with the user's operating system and compiling the software if necessary. Detailed installation instructions are provided on the GitHub page, ensuring that users can get DIAMOND up and running with minimal hassle.

Quick Start

Once DIAMOND is installed, users can quickly begin aligning sequences by preparing their input data in the form of DNA reads or protein sequences and a reference protein database. The tool comes with a set of straightforward commands that allow users to perform alignments, generate reports, and customize the sensitivity and specificity of the search.

A typical quick start command might involve specifying the input file containing the sequences to be aligned, the reference database, and the desired output file for the alignment results. Users can also select between fast and sensitive modes depending on their specific needs and the nature of their data.

Code Examples Of Popular Commands

Here are five popular commands that users of DIAMOND might find useful:

  1. Basic DIAMOND alignment command:

    diamond blastx -d nr -q reads.fna -o matches.m8

    This command aligns DNA sequences from reads.fna against the NR database and outputs the results in the m8 format to matches.m8.

  2. Using sensitive mode:

    diamond blastx --sensitive -d nr -q reads.fna -o matches.m8

    This command runs DIAMOND in sensitive mode for more comprehensive alignment results.

  3. Filtering by e-value:

    diamond blastx -d nr -q reads.fna -o matches.m8 -e 0.001

    This command filters the alignments to only include those with an e-value better than 0.001.

  4. Specifying the number of CPU threads:

    diamond blastx -d nr -q reads.fna -o matches.m8 -p 8

    This command specifies that DIAMOND should use 8 CPU threads to perform the alignment, which can speed up the process on multi-core systems.

  5. Generating a tabular report with more details:

    diamond blastx -d nr -q reads.fna -o matches.m8 -f 6 qseqid sseqid pident length mismatch gapopen qstart qend sstart send evalue bitscore

    This command generates a tabular report that includes detailed information about each alignment, such as query and subject IDs, percentage of identity, alignment length, mismatches, gap openings, query and subject start and end positions, e-value, and bit score.

These commands represent just a few examples of what can be done with DIAMOND. The tool's flexibility and range of options make it suitable for a wide array of sequence alignment tasks in bioinformatics research.

In conclusion, DIAMOND is a powerful and efficient tool for aligning DNA reads or protein sequences against large reference databases. Its speed and sensitivity make it an essential resource for researchers dealing with high-throughput sequence data. With easy installation and a variety of commands to customize the alignment process, DIAMOND is well-equipped to handle the demands of modern bioinformatics analysis.