IP Library › Granted Patent US 12,416,102
Granted Patent B2
US 12,416,102 · App. 17/397,775 · Granted Sep 16, 2025

Systems and methods for transfer of reagents between droplets

Inventors: Solongo B. Ziraldo (Pleasanton, CA); Geoffrey McDermott (Livermore, CA); Shea Thompson Lance (Livermore, CA)
Assignee: 10X GENOMICS, INC.
C40B50/06
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Quick Facts
Patent No.
US 12,416,102
App. No.
17/397,775
Granted
Sep 16, 2025
Kind
B2
Abstract

The disclosure provides systems and methods for droplet processing. For example, a method can include providing a first droplet and a second droplet. In some cases, the first droplet may have a first concentration of a reagent and the second droplet may have a second concentration of the reagent. The second droplet may comprise a bead or a biological particle. The method can also include subjecting the first droplet and the second droplet to conditions sufficient to transfer the reagent from the first droplet to the second droplet, thereby decreasing the first concentration in the first droplet and increasing the second concentration in the second droplet.

Claims (27)

1. A method for nucleic acid processing, comprising:

(a) generating a plurality of droplets by agitating:

(i) a first fluid volume comprising a population of beads and a population of analyte carriers, wherein a bead of the population of beads comprises a plurality of nucleic acid barcode molecules, and wherein an analyte carrier of the population of analyte carriers comprises a plurality of nucleic acid molecules, and

(ii) a second fluid volume immiscible with the first fluid volume,

wherein a first droplet of the plurality of droplets comprises the bead of the population of beads and the analyte carrier of the population of analyte carriers;

(b) subjecting the first droplet and a second droplet comprising a reagent to conditions sufficient to transfer the reagent to the first droplet from the second droplet via a mediator comprising an aggregate of surfactant molecules and the reagent, wherein the reagent is configured to release the plurality of nucleic acid molecules from the analyte carrier; and

(c) generating a barcoded nucleic acid molecule using a nucleic acid barcode molecule of the plurality of nucleic acid barcode molecules and a nucleic acid molecule of the plurality of nucleic acid molecules.

2. The method of claim 1 , wherein the population of analyte carriers is a population of cells or nuclei, and wherein the analyte carrier is a cell or a nucleus.

3. The method of claim 2 , wherein the reagent is a lysis agent.

4. The method of claim 3 , wherein the method comprises lysing the cell or the nucleus with the lysis agent in the first droplet.

5. The method of claim 4 , wherein the lysis agent is a surfactant.

6. The method of claim 4 , wherein the lysis agent is n-dodecyl-beta-D-maltoside (DBDM), sodium lauroyl sarcosinate, sodium dodecyl sulfate (SDS), t-Octylphenoxypolyethoxyethanol, or polysorbate 20.

7. The method of claim 4 , wherein the lysis agent is an enzyme.

8. The method of claim 1 , wherein the nucleic acid molecule is a messenger ribonucleic acid (mRNA).

9. The method of claim 8 , wherein the nucleic acid barcode molecule hybridizes to the mRNA.

10. The method of claim 9 , wherein generating the barcoded nucleic acid molecule comprises extending the nucleic acid barcode molecule in a reverse transcription reaction using the mRNA as template.

11. The method of claim 10 , wherein the method further comprises sequencing the barcoded nucleic acid molecule or a derivative thereof.

12. The method of claim 1 , wherein the mediator is a micelle.

13. The method of claim 1 , wherein the aggregate of surfactant molecules comprises a fluorosurfactant.

14. The method of claim 1 , wherein the second fluid volume comprises oil.

15. The method of claim 14 , wherein the oil is a fluorinated oil and the second fluid volume further comprises a fluorosurfactant.

16. The method of claim 1 , wherein the plurality of droplets comprises monodisperse droplets.

17. The method of claim 1 , wherein the agitating comprises vortexing.

18. The method of claim 1 , wherein the plurality of droplets is a first plurality of droplets, wherein the second droplet is a second droplet of a second plurality of droplets comprising the reagent, and wherein (b) comprises subjecting the first plurality of droplets and the second plurality of droplets to the conditions sufficient to transfer the reagent to the first droplet from the second droplet.

19. The method of claim 18 , wherein (b) comprises contacting the first plurality of droplets with the second plurality of droplets.

20. The method of claim 19 , wherein the first plurality of droplets is formed in a first emulsion, the second plurality of droplets is formed in a second emulsion, and wherein (b) comprises contacting the first emulsion with the second emulsion.

21. The method of claim 1 , wherein the population of analyte carriers is a population of cells or nuclei, and wherein the mediator is a micelle.

Assignments (1)
ASSIGNMENT OF ASSIGNOR'S INTEREST Recorded May 19, 2025
From: ZIRALDO, SOLONGO B.; MCDERMOTT, GEOFFREY; LANCE, SHEA THOMPSON
To: 10X GENOMICS, INC.
Reel/Frame 071158/0657 →
Continuity (4)
Continuation PCTUS2020017785 · Feb 11, 2020
Provisional Application 62934256 · Nov 12, 2019
Provisional Application 62804633 · Feb 12, 2019
Related Publication 20220025438A1 · Jan 27, 2022
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