US 12,467,088 B2
Probe-based analysis of nucleic acids and proteins
Katherine Pfeiffer, San Francisco, CA (US); Andrew Scott Kohlway, Dublin, CA (US); Andrew John Hill, Seattle, WA (US); and Paul Eugene Lund, San Leandro, CA (US)
Assigned to 10X GENOMICS, INC., Pleasanton, CA (US)
Filed by 10x Genomics, Inc., Pleasanton, CA (US)
Filed on Feb. 6, 2025, as Appl. No. 19/047,506.
Application 19/047,506 is a continuation of application No. 18/737,828, filed on Jun. 7, 2024, abandoned.
Application 18/737,828 is a continuation of application No. 18/593,799, filed on Mar. 1, 2024, abandoned.
Application 18/593,799 is a continuation of application No. 18/236,540, filed on Aug. 22, 2023, granted, now 11,952,626, issued on Apr. 9, 2024.
Application 18/236,540 is a continuation of application No. PCT/US2022/017377, filed on Feb. 22, 2022.
Claims priority of provisional application 63/251,446, filed on Oct. 1, 2021.
Claims priority of provisional application 63/235,487, filed on Aug. 20, 2021.
Claims priority of provisional application 63/196,834, filed on Jun. 4, 2021.
Claims priority of provisional application 63/152,709, filed on Feb. 23, 2021.
Prior Publication US 2025/0171845 A1, May 29, 2025
This patent is subject to a terminal disclaimer.
Int. Cl. C12Q 1/68 (2018.01); C12P 19/34 (2006.01); C12Q 1/6818 (2018.01); C12Q 1/6874 (2018.01)
CPC C12Q 1/6874 (2013.01) [C12Q 1/6818 (2013.01); C12Q 2600/16 (2013.01)] 30 Claims
OG exemplary drawing
 
1. A method of analyzing a sample, comprising:
(a) providing a plurality of cells, wherein:
(i) a cell of the plurality of cells comprises a first feature, wherein the first feature is an intracellular feature; and
(ii) the cell comprises a second feature, wherein the second feature is a cell surface feature;
(b) contacting the cell with:
(i) a first feature binding group, wherein the first feature binding group couples to the first feature in the cell, wherein the first feature binding group comprises a first oligonucleotide comprising a first feature nucleic acid sequence that identifies the first feature binding group; and
(ii) a second feature binding group, wherein the second feature binding group couples to the second feature in the cell, wherein the second feature binding group comprises a second oligonucleotide comprising a second feature nucleic acid sequence that identifies the second feature binding group;
(c) after (b), partitioning the plurality of cells and a plurality of barcode molecules into a plurality of partitions, wherein a partition of the plurality of partitions comprises the cell, a first barcode molecule comprising a first barcode sequence, and a second barcode molecule comprising a second barcode sequence;
(d) after (b), generating a first barcoded nucleic acid molecule using the first feature nucleic acid sequence and the first barcode molecule, wherein the first barcoded nucleic acid molecule comprises (i) the first feature nucleic acid sequence or reverse complement thereof, and (ii) the first barcode sequence or reverse complement thereof; and
(e) after (b), generating a second barcoded nucleic acid molecule using the second feature nucleic acid sequence and the second barcode molecule, wherein the second barcoded nucleic acid molecule comprises (i) the second feature nucleic acid sequence or reverse complement thereof, and (ii) the second barcode sequence or reverse complement thereof.