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US 2022/0177938 A1Published application
Polynucleotide synthesis method, kit and system
AssigneeOxford Nanopore Technologies PLC
InventorsJohn Milton, Sobia Nayyar, Jan Riedl, Ryosuke Ogaki
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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, as well as systems and kits for performing the synthesis and/or assembly methods.
Claims16
1-94. (canceled)
An in vitro method of synthesising a double-stranded polynucleotide having a predefined sequence, the method comprising performing repeating cycles of synthesis, wherein in each cycle:
- (A) a first strand of a double-stranded polynucleotide is extended by the addition of a first nucleotide of the predefined sequence by the action of a ligase enzyme in a blunt-ended ligation reaction;
- (B) the second strand of the double-stranded polynucleotide which is hybridized to the first strand is extended by the addition of a second nucleotide of the predefined sequence by a nucleotide transferase or polymerase enzyme;
- and
- (C) the double-stranded polynucleotide is then cleaved at a cleavage site;
- and
- wherein the first and second nucleotides of the predefined sequence of each cycle are retained in the double-stranded polynucleotide following cleavage.
3 more elements
Dependencies are inferred from the claim text.