High efficiency, small volume nucleic acid synthesis
AssigneesLife Technologies Corporation, Thermo Fisher Scientific Geneart GmbH
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).
Claims22
A multiwell plate for non-template directed synthesis of nucleic acid molecules, the plate comprising:
- (a) a bead located in each of a plurality of wells of the plate, and
- (b) an electrochemically generated acid being present in one or more well,
- wherein the bead is between 1.0 μm and 100 μm in diameter.
(canceled)
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 between 1×103 to 1×109 copies or in a volume of between 0.1 picoliters and 10 nanoliters;
- (b) combining the nucleic acid molecules generated in (a) to produce a pool;
- (c) joining some or all of the nucleic acid molecules present in the pool formed in (b) to form a plurality of larger nucleic acid molecules;
- (d) eliminating nucleic acid molecules which contain sequence errors from the plurality of larger nucleic acid molecules formed in (c) to produce an error corrected nucleic acid molecule pool;
- and
- (e) assembling the nucleic acid molecules in the error corrected nucleic acid molecule pool to form the assembled nucleic acid molecule.
-41. (canceled)
A method for producing a plurality of nucleic acid molecules, the method comprising:
- (a) synthesizing the plurality of nucleic acid molecules on separate beads in wells of a plate, wherein each well is configured to accommodate a single bead, wherein the volume of each well is between 0.1 picoliters and 10 nanoliters, wherein the wells comprise an electrode for the production of electrochemically generated acid, and wherein the nucleic acid molecules synthesized on each bead are designed to have the same nucleotide sequence;
- (b) selectively removing a plurality of beads from the wells, wherein the beads removed from the wells comprise a first set of individual nucleic acid molecules, wherein beads comprising nucleic acid molecules that are not part of the first set of individual nucleic acid molecules are not removed from their respective wells;
- and
- (c) collecting the beads selectively removed from wells in step (b) in a vessel.
Dependencies are inferred from the claim text.