Methods and Devices for Nucleic Acid Synthesis
AssigneeGen9, Inc.
InventorJoseph Jacobson
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Abstract
Disclosed are devices and methods to synthesize polynucleotides and libraries of polynucleotides such as libraries of oligonucleotides. In exemplary embodiments, the device includes a support having a plurality of features. Each feature contains a plurality of oligonucleotides. Within each feature, each of the plurality of oligonucleotides includes an identical predetermined subunit sequence of X nucleosides and a degenerate sequence of Y nucleosides. A predetermined combination of a subset of the features can be used to produce a polynucleotide having a predetermined sequence of Z nucleosides.
Claims20
A device for synthesizing at least one polynucleotide having a predetermined sequence, the device comprising:
- a solid support comprising a plurality of spots, wherein each of the plurality of spots comprises a plurality of oligonucleotides having a predetermined subunit sequence, wherein the plurality of oligonucleotides are covalently linked at the 3′ end via a plurality of binding sequences on the solid support;
- a microfluidic member for providing a droplet to a first spot having a plurality of first oligonucleotides having a first predetermined subunit sequence, wherein the droplet comprises one or more reagents that allow one or more of annealing, denaturing, chain extension, ligation, and digestion reaction to produce a first product comprising the first predetermined subunit sequence;
- and
- a member for advancing microfluidic communication between the first spot and a second spot, wherein the second spot has a second plurality of oligonucleotides having a second predetermined subunit sequence, whereby allowing one or more of annealing, denaturing, chain extension, ligation, and digestion reaction at the second spot to produce a second product comprising the first and the second predetermined subunit sequences.
A method for synthesizing at least one polynucleotide having a predetermined sequence, the method comprising:
- (a) providing a solid support having a plurality of spots thereon, wherein each of the plurality of spots comprises a plurality of oligonucleotides having a predetermined subunit sequence, wherein the plurality of oligonucleotides is covalently linked at the 3′ end via a plurality of binding sequences to the solid support;
- (b) providing a droplet to a first spot comprising a first plurality of oligonucleotides having a first predetermined subunit sequence, wherein the droplet comprises one or more reagents that allow one or more of annealing, denaturing, chain extension, ligation, and digestion reaction to produce a first product comprising the first predetermined subunit sequence;
- and
- (c) advancing microfluidic communication between the first spot and a second spot having a second plurality of oligonucleotides having a second predetermined subunit sequence, thereby allowing one or more of annealing, denaturing, chain extension, ligation, and digestion reaction at the second spot to produce a second product comprising the first and the second predetermined subunit sequences.
A device for synthesizing at least one polynucleotide having a predetermined sequence, the device comprising:
- a support having a plurality of features, each feature comprising a plurality of different oligonucleotides immobilized via their 3′ end, wherein each plurality of oligonucleotides has a different predetermined subunit sequence of X nucleosides at its 5′ end and has a degenerate sequence of Y nucleosides at its 3′ end;
- wherein the device has at least 4X different features,
- wherein a subset of the plurality of the features comprises oligonucleotides having different predetermined subunit sequences that together represent a polynucleotide having a predetermined sequence of Z nucleosides
Dependencies are inferred from the claim text.