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SEGUID v2: Extending SEGUID checksums for circular, linear, single- and double-stranded biological sequences
Humberto Pereira1, Paulo César Silva1, M Wayne Davis2
1Center of Molecular and Environmental Biology Engineering, University of Minho, Campus de Gualtar, Braga, Portugal.
Plos One
|August 7, 2026
Summary
SEGUIDv2 generates unique checksums for various DNA and RNA sequences, including circular and double-stranded types. This advancement improves data integrity and simplifies sequence identification in bioinformatics research.
Area of Science:
- Bioinformatics
- Molecular Biology
- Synthetic Biology
Background:
- Synthetic biology relies on combining DNA fragments for research and applications.
- Cryptographic checksums ensure data integrity for DNA sequences, aiding validation and annotation.
- Existing checksum methods like SEGUID are limited to specific DNA types (protein, ssDNA).
Purpose of the Study:
- To extend the SEGUID checksum algorithm to accommodate circular DNA and double-stranded DNA (dsDNA).
- To develop a universal checksum solution for linear and circular, single-stranded (ssDNA) and double-stranded (dsDNA) nucleic acid sequences.
- To address the limitations of the original SEGUID for handling diverse DNA structures.
Main Methods:
- Developed SEGUIDv2, an enhanced algorithm building upon the original SEGUID.
- Implemented SEGUIDv2 to generate orientation and rotation invariant checksums.
- Utilized Base64url encoding for SHA-1 hashes to ensure URL and filename compatibility.
Main Results:
- SEGUIDv2 successfully generates unique checksums for linear, circular, ssDNA, dsDNA, and RNA sequences.
- Checksums are invariant to sequence orientation and rotation.
- Base64url encoding enhances the usability of SEGUIDv2 checksums across different platforms and applications.
Conclusions:
- SEGUIDv2 provides a robust solution for sequence data integrity across various nucleic acid types.
- The algorithm's compatibility with filenames and URLs minimizes integration friction.
- Readily available implementations in major programming languages facilitate widespread adoption.
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