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Methods and systems for processing polynucleotides

DC
  • US 10,273,541 B2
  • Filed: 08/01/2018
  • Issued: 04/30/2019
  • Est. Priority Date: 08/14/2012
  • Status: Active Grant
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 microwell array comprising a plurality of wells, wherein a well of said plurality of wells comprises said 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 well 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, said released mRNA molecules attach to said nucleic acid barcode molecules;

    (c) subjecting said released 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 or a complement thereof; 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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