Systems and methods for processing-nucleic acid molecules from a single cell using sequential co-partitioning and composite barcodes
Assignee10x Genomics, Inc.
InventorsJessica Michele Terry, Tobias Daniel Wheeler
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Abstract
Provided herein are methods, compositions, and systems for multiplexed analysis of individual cells or cell populations. Cells encapsulated in beads and/or biomolecules are sequentially co-partitioned, allowing for analysis of two different types of biomolecules (e.g., RNA and DNA). The present invention leverages different polymer dissociation mechanisms, accompanied with barcoding of biomolecules (e.g., nucleic acid molecules) for multiplexed measurements in single cells. Sequential co-partitioning and barcode technology enables identification and quantitation of DNA and RNA from single cells.
Claims21
A method for barcoding nucleic acid molecules of a cell, comprising:
- (a) co-partitioning a cell bead and a first bead in a first partition, wherein said cell bead is generated from said cell, wherein said cell comprises a first nucleic acid molecule and a second nucleic acid molecule, wherein said first nucleic acid molecule comprises a different nucleic acid sequence than said second nucleic acid molecule, and wherein said first bead comprises a first set of barcode molecules each comprising a first nucleic acid barcode sequence;
- (b) subjecting said first partition to a first condition sufficient to permit (i) said first nucleic acid molecule to couple to a first barcode molecule of said first set of barcode molecules to generate a first barcoded nucleic acid molecule comprising said first nucleic acid barcode sequence, and (ii) a second barcode molecule of said first set of barcode molecules to associate with said cell bead;
- (c) co-partitioning said cell bead associated with said second barcode molecule or derivative thereof and a second bead in a second partition, wherein said second bead comprises a second set of barcode molecules each comprising a second nucleic acid barcode sequence;
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- (d) subjecting said second partition to a second condition sufficient to permit (i) said second nucleic acid molecule to couple to a first barcode molecule of said second set of barcode molecules to generate a second barcoded nucleic acid molecule comprising said second nucleic acid barcode sequence, and (ii) a second barcode molecule of said second set of barcode molecules to couple to said second barcode molecule to generate a composite barcode molecule comprising said first nucleic acid barcode sequence and said second nucleic acid barcode sequence.
Dependencies are inferred from the claim text.