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US 12,595,500 B2Granted
High efficiency, small volume nucleic acid synthesis
AssigneeLife Technologies Corporation
InventorsTodd Peterson, Axel Trefzer, Thomas Poehmerer
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Abstract
The disclosure generally relates to compositions and methods for the production of nucleic acid molecules. In some aspects, the invention allows for the microscale generation of nucleic acid molecules, optionally followed by assembly of these nucleic acid molecules into larger molecules. In some aspects, the invention allows for efficient production of nucleic acid molecules (e.g., large nucleic acid molecules such as genomes).
Claims14
A method for the generation of an assembled nucleic acid molecule, the method comprising:
- (a) synthesizing a plurality of nucleic acid molecules, wherein each nucleic acid molecule is prepared in a well of a plate in an average amount of from 0.001 nanomoles to 1,000 nanomoles;
- (b) combining the nucleic acid molecules generated in (a) to produce a pool of single-stranded nucleic acid molecules;
- (c) generating double-stranded nucleic acid molecules from nucleic acid molecules present in the pool of single-stranded nucleic acid molecules formed in step (b) to generate a pool of double-stranded nucleic acid molecules;
- (d) eliminating nucleic acid molecules which contain sequence errors from the pool of double-stranded nucleic acid molecules formed in step (c) by mismatch endonuclease correction to produce an error corrected pool of double-stranded nucleic acid molecules;
- and
- (e) simultaneously assembling the nucleic acid molecules in the error corrected pool of double-stranded nucleic acid molecules and an acceptor nucleic acid molecule to form the assembled nucleic acid molecule.
3 more elements
Dependencies are inferred from the claim text.