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US 2022/0064628 A1Published application
Compositions and methods for synthetic gene assembly
AssigneeTwist Bioscience Corporation
InventorsEsteban Toro, Sebastian TREUSCH, Siyuan CHEN, Cheng-Hsien WU
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Abstract
Methods and compositions are provided for assembly of large nucleic acids where the assembled large nucleic acids lack internal sequence modifications made during the assembly process.
Claims17
A nucleic acid assembly method comprising:
- a) obtaining at least two double-stranded DNA fragments, wherein the at least two DNA fragments comprise ends that can selectively hybridize with one another;
- and
- b) contacting the at least two double-stranded DNA fragments with:
- (i) a DNA polymerase;
- (ii) a flap endonuclease;
- and
- (iii) a DNA ligase,
- under reaction conditions that promote hybridization of the at least two DNA fragments and support the activities of components (i), (ii) and (iii), to produce a product comprising an assembled nucleic acid comprising at least a portion of each of the at least two DNA fragments.
5 more elements
A nucleic acid assembly method comprising:
- a) de novo synthesizing at least two single-stranded DNA fragments;
- b) amplifying the at least two single-stranded DNA fragments to generate at least two double single-stranded DNA fragments, wherein the at least two DNA fragments comprise ends that can selectively hybridize with one another;
- and
- c) contacting the at least two double-stranded DNA fragments with:
- (i) a DNA polymerase;
- (ii) a flap endonuclease;
- and
- (iii) a DNA ligase,
- under reaction conditions that promote hybridization of the at least two DNA fragments and support the activities of components (i), (ii) and (iii), to produce a product comprising an assembled nucleic acid comprising at least a portion of each of the at least two DNA fragments.
6 more elements
Dependencies are inferred from the claim text.