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US 2024/0263353 A1Published application
High-throughput single-cell polyomics
AssigneeYale University
InventorsRong Fan, Burak Dura
Life of this patent
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Abstract
Provided herein, in some embodiments, are devices, systems and methods for high-throughput single-cell polyomics (e.g., genomic, epigenomic, proteomic and/or phenotypic profile) analyses.
Claims26
A polyomic multiplexing device, comprising:
- a substrate comprising X rows intersecting Y columns to form X*Y patches, wherein each of the X*Y patches comprises a unique nucleic acid barcode that is immobilized to the substrate and comprises a polyT sequence,
- wherein each row comprises a different subset of barcoded nucleic acid strands of a first set of nucleic acid strands, and each column comprises a different subset of barcoded nucleic acid strands of a second set of nucleic acid strands, and
- wherein the nucleic acid strands of the first set are bound to nucleic acid strands of the second set to form a unique nucleic acid barcode.
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A polyomic multiplexing device, comprising:
- a microwell array comprising at least 20 microwells, wherein each microwell of the array comprises a molecular barcode specific to a single microwell, and wherein each molecular barcode comprises (a) a nucleic acid barcode that comprises a polyT sequence and (b) at least one antibody.
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A method of producing a barcoded array, comprising:
- (a) flow patterning and immobilizing onto a surface of a substrate a first set of barcoded nucleic acid strands of a first solution to produce columns that are parallel to and space apart relative to each other, wherein each column comprises X patches of barcoded nucleic acid strands of the first set, wherein the patches within each column are spaced apart relative to each other, wherein each column comprises a different subset of barcoded nucleic acid strands, and wherein X is a number greater than 2;
- (b) flow patterning and immobilizing onto the surface of the substrate a second set of barcoded nucleic acid strands of a second solution to produce rows that are parallel to and space apart relative to each other, wherein each row comprises Y patches of barcoded nucleic acid strands of the second set, wherein the patches within each row are spaced apart relative to each other, wherein each row comprises a different subset of barcoded nucleic acid strands, wherein the rows are perpendicular relative to the columns, and wherein Y is a number greater than 2,
- thereby producing a X*Y array of patches, each patch comprising (i) a subset of barcoded nucleic acid strands of the first set bound to (ii) a subset of barcoded nucleic acid strands of the second set to form a unique nucleic acid barcode.
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Dependencies are inferred from the claim text.