Patent ID: 8716467
Filing Date: 2014-05-06
Classification: C12M,C12N,C12P

Abstract:
1. 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, each plurality of oligonucleotides having a different predetermined subunit sequence at its 5′ end and a degenerate binding sequence at its 3′ end, wherein the plurality of oligonucleotides is covalently linked at the 3′ end via a plurality of binding sequences to the solid support and wherein the predetermined subunit sequences together comprise the at least one polynucleotide; (b) providing a droplet to a first predetermined spot, the first spot comprising a first plurality of oligonucleotides having a degenerate binding sequence and a first predetermined subunit sequence, wherein the droplet comprises a sequence capable of annealing to the degenerate sequence of the first plurality of oligonucleotides and one or more reagents that allow one or more of annealing, denaturing, chain extension, ligation, and digestion reaction to produce a first extension product comprising the first predetermined subunit sequence; (c) advancing a droplet comprising the first extension product from the first spot to a predetermined second spot, the second spot having a second plurality of oligonucleotides comprising a degenerate binding sequence and a second predetermined subunit sequence different than the first predetermined sequence; and (d) providing one or more reagents thereby allowing one or more of annealing, denaturing, chain extension, ligation, and digestion reaction at the second spot to produce a second extension product comprising the first and the second predetermined subunit sequences.