Sequencing by Expansion (SBX) - a novel, high-throughput single-molecule sequencing technology

Area of Science:

• Biotechnology
• Genomics
• Molecular Biology

Background:

• High-throughput sequencing advances drive biological discovery and clinical applications.
• Nanopore sequencing offers potential for scale and cost reduction but faces signal-to-noise limitations.
• Current nanopore methods struggle with nucleobase resolution, hindering accuracy and market adoption.

Purpose of the Study:

• To introduce Sequencing by Expansion (SBX), a novel single-molecule sequencing technology.
• To overcome the signal-to-noise and molecular distinction limitations of existing nanopore sequencing.
• To enable facile, high-accuracy nanopore sequencing through a new biochemical conversion process.

Main Methods:

• Encoding target nucleic acid sequences into Xpandomers, a surrogate polymer.
• Engineering Xpandomers for over 50-fold expansion compared to parent DNA templates.
• Utilizing high signal-to-noise reporter codes for enhanced nanopore detection.

Main Results:

• Demonstrated the performance and feasibility of the Sequencing by Expansion (SBX) technology.
• Developed specialized molecular structures, chemistries, and enzymes for SBX.
• Achieved high-accuracy nanopore sequencing through the Xpandomer conversion process.

Conclusions:

• SBX technology effectively addresses fundamental limitations in nanopore sequencing.
• The innovative molecular and systems engineering in SBX offers a transformative approach.
• SBX is poised to meet the demands of current and future sequencing applications.