MASSIVELY PARALLEL SINGLE CELL ANALYSIS
First Claim
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1. A method for processing messenger ribonucleic acid (mRNA) molecules from a single cell, comprising:
- (a) partitioning a plurality of cells and a plurality of beads in a plurality of partitions, wherein a partition of said plurality of partitions comprises a single cell from said plurality of cells and a single bead from said plurality of beads, and wherein said single bead comprises nucleic acid barcode molecules each comprising a common barcode sequence;
(b) in said partition comprising said single cell and said single bead, releasing messenger ribonucleic acid (mRNA) molecules from said single cell, wherein upon release from said single cell, whereby said mRNA molecules attach to said nucleic acid barcode molecules;
(c) subjecting said mRNA molecules attached to said nucleic acid barcode molecules to reverse transcription to yield complementary deoxyribonucleic acid (cDNA) molecules each comprising said common barcode sequence; and
(d) subjecting said cDNA molecules to one or more reactions to generate a set of nucleic acid molecules for nucleic acid sequencing.
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Abstract
The disclosure provides for methods, compositions, and kits for multiplex nucleic acid analysis of single cells. The methods, compositions and systems may be used for massively parallel single cell sequencing. The methods, compositions and systems may be used to analyze thousands of cells concurrently. The thousands of cells may comprise a mixed population of cells (e.g., cells of different types or subtypes, different sizes).
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20 Claims
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1. A method for processing messenger ribonucleic acid (mRNA) molecules from a single cell, comprising:
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(a) partitioning a plurality of cells and a plurality of beads in a plurality of partitions, wherein a partition of said plurality of partitions comprises a single cell from said plurality of cells and a single bead from said plurality of beads, and wherein said single bead comprises nucleic acid barcode molecules each comprising a common barcode sequence; (b) in said partition comprising said single cell and said single bead, releasing messenger ribonucleic acid (mRNA) molecules from said single cell, wherein upon release from said single cell, whereby said mRNA molecules attach to said nucleic acid barcode molecules; (c) subjecting said mRNA molecules attached to said nucleic acid barcode molecules to reverse transcription to yield complementary deoxyribonucleic acid (cDNA) molecules each comprising said common barcode sequence; and (d) subjecting said cDNA molecules to one or more reactions to generate a set of nucleic acid molecules for nucleic acid sequencing. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20)
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Specification