Methods and systems for processing polynucleotides
First Claim
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1. A method for identifying a protein as originating from a single cell, comprising:
- (a) exposing said protein of said single cell to one or more antibodies such that an antibody of said one or more antibodies that is specific to said protein couples to said protein, wherein said antibody is conjugated to a nucleic acid antibody barcode molecule comprising an antibody barcode sequence;
(b) co-partitioning said single cell comprising said protein coupled to said antibody with a single bead in a partition, wherein said single bead comprises a plurality of nucleic acid cell barcode molecules each comprising a cell barcode sequence;
(c) performing one or more reactions to generate, from said nucleic acid antibody barcode molecule and a nucleic acid cell barcode molecule from said plurality of nucleic acid cell barcode molecules, a synthesized nucleic acid molecule comprising (i) said antibody barcode sequence or a complement thereof, and (ii) said cell barcode sequence or a complement thereof;
(d) using a sequencer to sequence at least a portion of said synthesized nucleic acid molecule or a derivative thereof, to identify said antibody barcode sequence and said cell barcode sequence, and(e) using said antibody barcode sequence and said cell barcode sequence to identify said protein as originating from said single cell.
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Abstract
The present disclosure provides compositions, methods, systems, and devices for polynucleotide processing. Such polynucleotide processing may be useful for a variety of applications, including polynucleotide sequencing.
584 Citations
38 Claims
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1. A method for identifying a protein as originating from a single cell, comprising:
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(a) exposing said protein of said single cell to one or more antibodies such that an antibody of said one or more antibodies that is specific to said protein couples to said protein, wherein said antibody is conjugated to a nucleic acid antibody barcode molecule comprising an antibody barcode sequence; (b) co-partitioning said single cell comprising said protein coupled to said antibody with a single bead in a partition, wherein said single bead comprises a plurality of nucleic acid cell barcode molecules each comprising a cell barcode sequence; (c) performing one or more reactions to generate, from said nucleic acid antibody barcode molecule and a nucleic acid cell barcode molecule from said plurality of nucleic acid cell barcode molecules, a synthesized nucleic acid molecule comprising (i) said antibody barcode sequence or a complement thereof, and (ii) said cell barcode sequence or a complement thereof; (d) using a sequencer to sequence at least a portion of said synthesized nucleic acid molecule or a derivative thereof, to identify said antibody barcode sequence and said cell barcode sequence, and (e) using said antibody barcode sequence and said cell barcode sequence to identify said protein as originating from said single cell. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38)
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Specification