Polynucleotide synthesis method, kit and system

The method of cyclic ligation and cleavage of blunt-ended polynucleotides allows for the efficient synthesis of double-stranded polynucleotides with predefined sequences, overcoming template and condition limitations in existing methods.

WO2025003669A1PCT designated stage expired Publication Date: 2025-01-02OXFORD NANOPORE TECH LTD
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Patent Information

Application Number
PCT/GB2024/051643
Authority / Receiving Office
WO · WO
Patent Type
Applications
Current Assignee / Owner
Priority Date
2023-06-27
Filing Date
2024-06-27
Publication Date
2025-01-02

AI Technical Summary

Technical Problem

Current methods for synthesizing polynucleotides, such as phosphoramidite chemistry and polymerase synthesis, are limited in length and require pre-existing templates, while template-independent methods face issues like nucleotide bias and the need for reversible blocking groups, restricting the synthesis of double-stranded DNA de novo.

Method used

A method involving cycles of single-stranded ligation and cleavage using blunt-ended donor and acceptor polynucleotides, where a polynucleotide payload is ligated and then cleaved to extend the acceptor, allowing stepwise synthesis of double-stranded polynucleotides without extreme conditions or templates, under mild aqueous conditions.

Benefits of technology

Enables the de novo synthesis of double-stranded polynucleotides with predefined sequences, avoiding template dependency and extreme conditions, and improving synthesis efficiency by minimizing reagent loss and random ligation.

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Abstract

The invention relates to new methods for synthesising polynucleotide molecules according to a predefined nucleotide sequence. The invention also relates to methods for the assembly of synthetic polynucleotides following synthesis.
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Citation Information

Patent Citations

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  • Am-EWOD device and method of driving with variable voltage ac driving

    US20140202863A1

  • Methods and apparatus for synthesizing nucleic acids

    US20140363852A1

  • Methods and apparatus for synthesizing nucleic acid

    US20160046973A1

  • Modified template-independent enzymes for polydeoxynucleotide synthesis

    US20160108382A1

Cited By

  • Polynucleotide synthesis methods

    WO2026132201A1