US 11,732,300 B2
Increasing efficiency of spatial analysis in a biological sample
Felice Alessio Bava, Pleasanton, CA (US)
Assigned to 10x Genomics, Inc., Pleasanton, CA (US)
Filed by 10x Genomics, Inc., Pleasanton, CA (US)
Filed on Feb. 4, 2021, as Appl. No. 17/167,559.
Claims priority of provisional application 62/970,633, filed on Feb. 5, 2020.
Prior Publication US 2021/0238675 A1, Aug. 5, 2021
Int. Cl. C12Q 1/6874 (2018.01); C12N 15/10 (2006.01)
CPC C12Q 1/6874 (2013.01) [C12N 15/1065 (2013.01)] 17 Claims
OG exemplary drawing
 
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.