US 12,264,411 B2
Methods and systems for analysis
Zahra Kamila Belhocine, Fremont, CA (US); Rajiv Bharadwaj, Pleasanton, CA (US); Christopher Hindson, Pleasanton, CA (US); Michael Schnall-Levin, San Francisco, CA (US); Bill Kengli Lin, Pleasanton, CA (US); Anthony Makarewicz, Livermore, CA (US); Pranav Patel, Fremont, CA (US); Katherine Pfeiffer, Berkeley, CA (US); Andrew D. Price, Hayward, CA (US); Mohammad Rahimi Lenji, Pleasanton, CA (US); Tobias Wheeler, Alameda, CA (US); and Yifeng Yin, Elk Grove, CA (US)
Assigned to 10X GENOMICS, INC., Pleasanton, CA (US)
Filed by 10x Genomics, Inc., Pleasanton, CA (US)
Filed on Mar. 1, 2024, as Appl. No. 18/593,788.
Application 18/593,788 is a continuation of application No. 17/517,282, filed on Nov. 2, 2021.
Application 17/517,282 is a continuation of application No. 17/208,369, filed on Mar. 22, 2021, granted, now 11,193,122, issued on Dec. 7, 2021.
Application 17/208,369 is a continuation of application No. 16/419,461, filed on May 22, 2019, abandoned.
Application 16/419,461 is a continuation of application No. 15/887,947, filed on Feb. 2, 2018, granted, now 10,428,326, issued on Oct. 1, 2019.
Application 15/887,947 is a continuation of application No. PCT/US2018/016019, filed on Jan. 30, 2018.
Claims priority of provisional application 62/452,261, filed on Jan. 30, 2017.
Claims priority of provisional application 62/500,943, filed on May 3, 2017.
Claims priority of provisional application 62/570,783, filed on Oct. 11, 2017.
Prior Publication US 2024/0254658 A1, Aug. 1, 2024
Int. Cl. C40B 30/04 (2006.01); C07K 14/74 (2006.01); C12N 15/10 (2006.01); C12N 15/11 (2006.01); C12N 15/85 (2006.01); C12Q 1/6804 (2018.01); C12Q 1/6806 (2018.01); C12Q 1/6818 (2018.01); C12Q 1/6827 (2018.01); C12Q 1/6881 (2018.01); C40B 50/06 (2006.01); G01N 33/50 (2006.01); G01N 33/53 (2006.01); G01N 33/532 (2006.01); G01N 33/543 (2006.01); G01N 33/548 (2006.01); G01N 33/569 (2006.01); G01N 33/58 (2006.01); C40B 70/00 (2006.01)
CPC C40B 30/04 (2013.01) [C07K 14/70539 (2013.01); C12N 15/1037 (2013.01); C12N 15/1055 (2013.01); C12N 15/1065 (2013.01); C12N 15/1075 (2013.01); C12N 15/11 (2013.01); C12N 15/85 (2013.01); C12Q 1/6804 (2013.01); C12Q 1/6806 (2013.01); C12Q 1/6818 (2013.01); C12Q 1/6827 (2013.01); C12Q 1/6881 (2013.01); G01N 33/5032 (2013.01); G01N 33/505 (2013.01); G01N 33/5304 (2013.01); G01N 33/5306 (2013.01); G01N 33/5308 (2013.01); G01N 33/532 (2013.01); G01N 33/54306 (2013.01); G01N 33/54366 (2013.01); G01N 33/548 (2013.01); G01N 33/56977 (2013.01); G01N 33/58 (2013.01); C12N 2310/20 (2017.05); C12N 2320/10 (2013.01); C12Q 2537/164 (2013.01); C12Q 2563/179 (2013.01); C12Q 2563/185 (2013.01); C12Q 2565/1015 (2013.01); C40B 50/06 (2013.01); C40B 70/00 (2013.01)] 20 Claims
 
1. A method for processing or analyzing a single cell or nucleus, the method comprising:
(a) forming a partition, comprising:
(i) providing (1) an aqueous first liquid comprising a single cell or nucleus, (2) an aqueous second liquid comprising one or more polymeric or gel precursors, and (3) a third liquid that is immiscible with the first liquid and the second liquid;
(ii) forming a droplet comprising an aqueous two-phase system surrounded by the third liquid, wherein the aqueous first liquid and the aqueous second liquid form an inner phase and an outer phase, respectively, of the aqueous two-phase system;
(iii) providing a stimulus to the outer phase to generate a gel or polymer shell encapsulating the inner phase comprising the single cell or nucleus to thereby form the partition, wherein the gel or polymer shell is diffusively permeable to one or more reagents; and
(b) introducing the one or more reagents to the inner phase of the partition comprising the single cell or nucleus via diffusion through the gel or polymer shell encapsulating the inner phase comprising the single cell or nucleus.