Methods and devices for nucleic acid synthesis
First Claim
1. A method for synthesizing a target polynucleotide having a predetermined sequence, the method comprising(a) providing a solid support having a first feature and a second feature thereon, the first feature having a first plurality of immobilized oligonucleotides each comprising a first predetermined subunit sequence and a first degenerate binding sequence, the second feature having a second plurality of immobilized oligonucleotides each comprising a second predetermined subunit sequence and a second degenerate binding sequence, wherein the first predetermined subunit sequence and the second predetermined subunit sequence together comprise a target polynucleotide;
- (b) generating, in a chain extension reaction using an input oligonucleotide as primer that anneals with the first degenerate binding sequence, a first product comprising the first predetermined subunit sequence at the first feature;
(c) advancing, via microfluidic communication, the first product to the second feature; and
(d) generating, in a chain extension reaction using the first product as primer that anneals with the second degenerate binding sequence, a second product comprising the first and the second predetermined subunit sequences at the second feature.
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Accused Products
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.
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Citations
18 Claims
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1. A method for synthesizing a target polynucleotide having a predetermined sequence, the method comprising
(a) providing a solid support having a first feature and a second feature thereon, the first feature having a first plurality of immobilized oligonucleotides each comprising a first predetermined subunit sequence and a first degenerate binding sequence, the second feature having a second plurality of immobilized oligonucleotides each comprising a second predetermined subunit sequence and a second degenerate binding sequence, wherein the first predetermined subunit sequence and the second predetermined subunit sequence together comprise a target polynucleotide; -
(b) generating, in a chain extension reaction using an input oligonucleotide as primer that anneals with the first degenerate binding sequence, a first product comprising the first predetermined subunit sequence at the first feature; (c) advancing, via microfluidic communication, the first product to the second feature; and (d) generating, in a chain extension reaction using the first product as primer that anneals with the second degenerate binding sequence, a second product comprising the first and the second predetermined subunit sequences at the second feature. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18)
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Specification