Methods and Devices for Nucleic Acid Synthesis
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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.
70 Citations
47 Claims
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1-20. -20. (canceled)
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21. A device for synthesizing at least one polynucleotide having a predetermined sequence, the device comprising:
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a solid support; a plurality of features onto the solid support, a plurality of different oligonucleotide sequences covalently linked via a plurality of binding sequences at the plurality of features, each plurality of different oligonucleotide sequences having a different binding sequence having a length N1 nucleosides and a different predetermined subunit sequence having a length N2 nucleosides; the plurality of binding sequences comprising up to 4N1 different binding sequences and the plurality of features comprising up to 4N2 features. - View Dependent Claims (29, 30)
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22-28. -28. (canceled)
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31. A method for synthesizing a polynucleotide having a predetermined sequence, the method comprising
(a) providing a solid support having a plurality of features thereon, the plurality of features comprising 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 each plurality of oligonucleotides is immobilized at the 3′
end to the solid support;(b) generating a first product comprising the first predetermined subunit sequence at a first feature; (c) advancing the first product to a second feature comprising the second predetermined subunit sequence, wherein the second predetermined subunit sequence is different from the first predetermined subunit sequence; and (d) generating a second product comprising the first and the second predetermined subunit sequences at the second feature. - View Dependent Claims (32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44)
- end and a degenerate binding sequence at its 3′
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45. A composition comprising a plurality of oligonucleotides, each oligonucleotide having a different predetermined subunit sequence at its 5′
- end and a different degenerate binding sequence at its 3′
end, wherein the degenerate sequences have a length of N1 nucleosides, and each degenerate sequence is one of 4N1 possible sequences. - View Dependent Claims (46, 47)
- end and a different degenerate binding sequence at its 3′
Specification