CPC C12Q 1/6874 (2013.01) [C12N 15/1065 (2013.01)] | 17 Claims |
1. A method of determining the location of an analyte in a biological sample, the method comprising:
(a) contacting a biological sample with a substrate comprising a plurality of capture probes, wherein a capture probe in the plurality of capture probes comprises a capture domain and a spatial barcode;
(b) hybridizing the analyte to the capture domain, thereby generating a capture analyte; and
(c) contacting the captured analyte to a bridging oligonucleotide, wherein the bridging oligonucleotide comprises:
(i) a capture-probe-binding sequence, and
(ii) an analyte-binding sequence;
(d) extending the bridging oligonucleotide using the analyte as a template to generate an extended capture probe comprising (i) the analyte, or a complement thereof, and (ii) the spatial barcode or a complement thereof; and
(e) determining (i) all or a part of the sequence of the analyte, or a complement thereof, and (ii) the sequence of the spatial barcode, or a complement thereof, and using the determined sequence of (i) and (ii) to determine the location of the analyte in the biological sample.
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