Methods and compositions for integrated in situ spatial assay
Assignee10x Genomics, Inc.
InventorsFelice Alessio BAVA, Zachary BENT
Life of this patent
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Abstract
In some embodiments, provided herein is an integrated assay of a biological sample comprising an in situ assay module and a spatial assay module. The in situ assay comprises analyzing binding between nucleic acid probes and a first analyte at a spatial location of the biological sample. In some embodiments, the method further comprises providing conditions to allow spatially barcoded capture agents to capture a second analyte for analysis in the spatial assay module.
Claims39
A method of analyzing a biological sample, comprising:
- (a) contacting a biological sample on a substrate with one or more nucleic acid probes that directly or indirectly hybridize to a first target nucleic acid in the biological sample,
- wherein the substrate comprises a plurality of capture agents directly or indirectly immobilized thereon, wherein a capture agent of the plurality of capture agents comprises (i) a capture domain capable of capturing a nucleic acid and (ii) a spatial barcode corresponding to the position of the capture agent on the substrate;
- (b) detecting the one or more nucleic acid probes at a spatial location of the biological sample;
- (c) providing conditions to allow the capture agents to directly or indirectly capture a second target nucleic acid;
- and
- (d) generating a spatially labeled polynucleotide comprising (i) a sequence of the second target nucleic acid or complement thereof and (ii) a sequence of the spatial barcode or complement thereof.
A method of analyzing a biological sample, comprising:
- (a) contacting a biological sample on a first substrate with one or more nucleic acid probes that directly or indirectly hybridize to a first target nucleic acid in the biological sample;
- (b) detecting the one or more nucleic acid probes at a spatial location of the biological sample;
- (c) providing a plurality of capture agents to directly or indirectly capture a second target nucleic acid, wherein the plurality of capture agents are joined directly or indirectly to a second substrate and a capture agent of the plurality of capture agents comprises:
- (i) a capture domain capable of capturing the second target nucleic acid, and
- (ii) a spatial barcode;
- and
- (d) generating a spatially labeled polynucleotide comprising (i) a sequence of the second target nucleic acid or complement thereof and (ii) a sequence of the spatial barcode or complement thereof.
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Dependencies are inferred from the claim text.