Method for sequential sequencing nucleic acids
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, 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;
performing successive cycles of a base-by-base sequencing for the target polynucleotides immobilized on the microparticles in the flow cell; and
obtaining data that provides sequence information about the plurality of target polynucleotides based on the successive cycles of the base-by-base sequencing.
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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.
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Citations
20 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, 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; performing successive cycles of a base-by-base sequencing for the target polynucleotides immobilized on the microparticles in the flow cell; and obtaining data that provides sequence information about the plurality of target polynucleotides based on the successive cycles of the base-by-base sequencing. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20)
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Specification