Methods and systems for sample preparation and analysis

Patent metadata

Field	Value
Publication number	US-12473585-B2
Application number	US-202318097989-A
Country	US
Kind code	B2
Filing date	Jan 17, 2023
Priority date	Oct 27, 2017
Publication date	Nov 18, 2025
Grant date	Nov 18, 2025

Abstract

The present disclosure provides methods and systems for sample preparation and/or analysis. Samples may be cells, or may be derived from one or more cells. Sample preparation may comprise conducting one or more reactions on a target. Such reactions may be conducted in one or more partitions. One or more reactions may be performed in one or more successive operations.

First claim

What is claimed is:

1 . A method for sample preparation or analysis, comprising:
(a) providing a plurality of partitions, wherein a given partition of the plurality of partitions comprises a cell and a first bead, wherein the first bead comprises one or more reagents and is disruptable; (b) disrupting the first bead in the given partition to produce one or more precursors and release the one or more reagents; and (c) using the one or more precursors to generate a second bead, which second bead comprises the cell and the one or more reagents, wherein the second bead is within the given partition.

2 . The method of claim 1 , further comprising performing one or more reactions on the cell using the one or more reagents.

3 . The method of claim 2 , wherein the one or more reactions comprise cell lysis.

4 . The method of claim 2 , wherein the one or more reactions comprise an addition of a barcode oligonucleotide to a nucleic acid molecule in the cell.

5 . The method of claim 2 , wherein the one or more reactions comprise tagmentation.

6 . The method of claim 2 , wherein the one or more reactions comprise a reverse transcription reaction.

7 . The method of claim 2 , wherein the one or more reactions comprise a nucleic acid modification.

8 . The method of claim 7 , wherein the nucleic acid modification is a deoxyribonucleic acid modification.

9 . The method of claim 7 , wherein the nucleic acid modification is a ribonucleic acid modification.

10 . The method of claim 2 , wherein the one or more reactions comprise nucleic acid amplification.

11 . The method of claim 10 , wherein the nucleic acid amplification is polymerase chain reaction.

12 . The method of claim 1 , wherein the plurality of partitions is a plurality of droplets.

13 . The method of claim 1 , wherein the plurality of partitions is a plurality of wells.

14 . The method of claim 1 , wherein the first bead is a gel bead.

15 . The method of claim 1 , wherein the second bead is a gel bead.

16 . The method of claim 1 , wherein the disrupting comprises application of a stimulus.

17 . The method of claim 16 , wherein the stimulus is a chemical stimulus, thermal stimulus, electromagnetic stimulus, or a combination thereof.

18 . The method of claim 1 , wherein the first bead is from a plurality of beads, and wherein the plurality of beads comprises nucleic acid barcode sequences that are different across the plurality of beads.

19 . A method for sample preparation or analysis, comprising:
(a) providing a plurality of partitions, wherein a given partition of the plurality of partitions comprises a cell and a first bead, wherein the first bead comprises one or more barcode molecules and is disruptable; (b) disrupting the first bead in the given partition to produce one or more precursors and release the one or more barcode molecules; (c) using the one or more precursors to generate a second bead, which second bead comprises the cell and the one or more barcode molecules, wherein the second bead is within the given partition; (d) lysing the cell in the second bead to generate cellular components; (e) disrupting the second bead; and (f) barcoding the cellular components using the one or more barcode molecules.

20 . A method for sample preparation or analysis, comprising:
(a) providing a plurality of partitions, wherein a given partition of the plurality of partitions comprises a cell, a first disruptable bead comprising one or more first molecular barcodes, and cellular growth media; (b) disrupting the first disruptable bead to produce one or more precursors and release the one or more first molecular barcodes in the given partition; (c) using the one or more precursors to generate a second bead, wherein the second bead comprises the cell and the one or more first molecular barcodes, wherein the second bead is within the given partition; (d) lysing the cell in the second bead, thereby generating cellular components; (e) partitioning the second bead in a second partition with a second disruptable bead comprising one or more second molecular barcodes; (f) disrupting the second bead to form second precursors, and disrupting the second disruptable bead; and (g) barcoding the cellular components using the one or more first molecular barcodes and the one or more second molecular barcodes.

Assignees

• 10X Genomics Inc

Inventors

• Belhocine Zahra Kamila
• Lee Josephine
• Meschi Francesca
• Montesclaros Luz
• Pfeiffer Katherine
• Price Andrew D
• Sapida Jerald
• Sukovich David
• Wheeler Tobias Daniel
• Yin Yifeng

Classifications

• B01L2200/0647

  Handling flowable solids, e.g. microscopic beads, cells, particles · CPC title

• C12N15/1075 Primary

  by coupling phenotype to genotype, not provided for in other groups of this subclass · CPC title

• B01L3/502784 Primary

  specially adapted for droplet or plug flow, e.g. digital microfluidics · CPC title

• C12Q1/6806 Primary

  Preparing nucleic acids for analysis, e.g. for polymerase chain reaction [PCR] assay (C12Q1/6804 takes precedence) · CPC title