System and apparatus for sequential processing of analytes
First Claim
1. A method of sequencing multiple target polynucleotides in parallel using a sequencing system, comprising:
- providing a flow chamber comprising a plurality of ridges within the flow chamber running parallel to the axis of the flow chamber, the flow chamber comprising multiple microparticles immobilized between the ridges, each microparticle of the multiple microparticles including a target polynucleotide to be sequenced immobilized on the microparticle, the average distance between centers of adjacent microparticles being less than two microparticle diameters, and the ridges being of a height less than a diameter of a microparticle and of a length greater than a plurality of said diameters;
providing an oligonucleotide through an inlet of the flow chamber;
ligating the oligonucleotide to a strand hybridized to a polynucleotide of the plurality of target polynucleotides when the oligonucleotide is complementary to a single-stranded portion of the polynucleotide;
obtaining data that provides information about the plurality of target polynucleotides based on the ligation of the oligonucleotides, the information produced being governed by at least a portion of a sequence of the plurality of target polynucleotides; and
cleaving at least a portion of the oligonucleotide that was ligated.
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Accused Products
Abstract
An apparatus and system are provided for simultaneously analyzing a plurality of analytes anchored to microparticles. Microparticles each having a uniform population of a single kind of analyte attached are disposed as a substantially immobilized planar array inside of a flow chamber where steps of an analytical process are carried out by delivering a sequence of processing reagents to the microparticles by a fluidic system under microprocessor control. In response to such process steps, an optical signal is generated at the surface of each microparticle which is characteristic of the interaction between the analyte carried by the microparticle and the delivered processing reagent. The plurality of analytes are simultaneously analyzed by collecting and recording images of the optical signals generated by all the microparticles in the planar array. A key feature of the invention is the correlation of the sequence of optical signals generated by each microparticle in the planar array during the analytical process.
226 Citations
29 Claims
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1. A method of sequencing multiple target polynucleotides in parallel using a sequencing system, comprising:
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providing a flow chamber comprising a plurality of ridges within the flow chamber running parallel to the axis of the flow chamber, the flow chamber comprising multiple microparticles immobilized between the ridges, each microparticle of the multiple microparticles including a target polynucleotide to be sequenced immobilized on the microparticle, the average distance between centers of adjacent microparticles being less than two microparticle diameters, and the ridges being of a height less than a diameter of a microparticle and of a length greater than a plurality of said diameters; providing an oligonucleotide through an inlet of the flow chamber; ligating the oligonucleotide to a strand hybridized to a polynucleotide of the plurality of target polynucleotides when the oligonucleotide is complementary to a single-stranded portion of the polynucleotide; obtaining data that provides information about the plurality of target polynucleotides based on the ligation of the oligonucleotides, the information produced being governed by at least a portion of a sequence of the plurality of target polynucleotides; and cleaving at least a portion of the oligonucleotide that was ligated. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26)
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27. A method of sequencing multiple target polynucleotides in parallel using a sequencing system, comprising:
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providing an oligonucleotide through an inlet of a flow chamber, said flow chamber comprising multiple microparticles immobilized between ridges within the flow chamber running parallel to the axis of the flow chamber, wherein said multiple microparticles include multiple target polynucleotides to be sequenced, wherein the average distance between centers of adjacent microparticles is less than two microparticle diameters, and wherein the ridges are of a height less than a diameter of a microparticle and of a length greater than a plurality of said diameters; and sequencing at least a portion of more than one of said multiple target polynucleotides. - View Dependent Claims (28, 29)
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Specification