US 12,305,239 B2
Analysis of nucleic acid sequences
Solongo B. Ziraldo, Pleasanton, CA (US); Geoffrey McDermott, Livermore, CA (US); and Shea Thompson Lance, Livermore, CA (US)
Assigned to 10X GENOMICS, INC., Pleasanton, CA (US)
Filed by 10x Genomics, Inc., Pleasanton, CA (US)
Filed on Nov. 18, 2024, as Appl. No. 18/951,533.
Application 18/951,533 is a continuation in part of application No. 17/397,775, filed on Aug. 9, 2021.
Application 17/397,775 is a continuation of application No. PCT/US2020/017785, filed on Feb. 11, 2020.
Claims priority of provisional application 62/934,256, filed on Nov. 12, 2019.
Claims priority of provisional application 62/804,633, filed on Feb. 12, 2019.
Prior Publication US 2025/0075272 A1, Mar. 6, 2025
This patent is subject to a terminal disclaimer.
Int. Cl. C12Q 1/6883 (2018.01); C12Q 1/6827 (2018.01); G16B 30/00 (2019.01); G16B 30/10 (2019.01); G16B 30/20 (2019.01)
CPC C12Q 1/6883 (2013.01) [C12Q 1/6827 (2013.01); G16B 30/00 (2019.02); G16B 30/10 (2019.02); G16B 30/20 (2019.02); C12Q 2535/122 (2013.01); C12Q 2600/156 (2013.01)] 30 Claims
 
1. A method comprising:
(a) generating a first plurality of droplets, wherein a first droplet of the first plurality of droplets comprises a lysis agent;
(b) partitioning a plurality of cells into a second plurality of droplets, wherein a second droplet of the second plurality of droplets comprises: i) a cell of the plurality of cells, and ii) a plurality of nucleic acid barcode molecules coupled to a particle;
(c) generating a micellized lysis agent from the first droplet, wherein the micellized lysis agent comprises a micelle comprising the lysis agent;
(d) delivering the lysis agent to the second droplet using the micellized lysis agent;
(e) lysing the cell within the second droplet with the lysis agent, thereby releasing a messenger RNA (mRNA) molecule from the cell into the second droplet;
(f) hybridizing the mRNA molecule to a nucleic acid barcode molecule of the plurality of nucleic acid barcode molecules; and
(g) generating a barcoded nucleic acid molecule using the mRNA molecule and the nucleic acid barcode molecule, wherein the barcoded nucleic acid molecule comprises: i) a sequence of the mRNA molecule or a complement thereof, and ii) a sequence of the nucleic acid barcode molecule or a complement thereof.