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US 2023/0323453 A1Published application
Enhancing specificity of analyte binding
OwnerMarlon Stoeckiusinventor
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Abstract
Methods for enhancing specificity of an analyte binding moiety or probe oligonucleotide to an analyte are provided herein. For example, methods provided herein include blocking a capture binding domain, thereby preventing hybridization to the capture domain of the capture probe affixed to a substrate. Further methods include releasing the block from the capture binding domain, thereby allowing the capture binding domain to specifically bind to the capture domain of the capture probe on the substrate.
Claims1
A method for identifying the location of an analyte in a biological sample comprising:
- (a) contacting the biological sample with a substrate comprising a plurality of capture probes comprising a spatial barcode and a capture domain;
- (b) binding an analyte-binding moiety to the analyte, wherein the analyte-binding moiety is associated with an oligonucleotide comprising an analyte-binding-moiety barcode and a capture binding domain that hybridizes to the capture domain of the capture probe, wherein a portion of the capture binding domain is hybridized to a blocking probe;
- (c) releasing the blocking probe from the capture binding domain and hybridizing the capture binding domain to the capture domain;
- and
- (d) determining (i) all or a part of the sequence of the oligonucleotide bound to the capture domain, or a complement thereof, and (ii) all or a part of the sequence of the spatial barcode, or a complement thereof, and using the determined sequence of (i) and (ii) to identify a location of the analyte in the biological sample.
2 more elements
Dependencies are inferred from the claim text.