Methods for analyzing nucleic acids from single cells

  • US 10,155,981 B2
  • Filed: 08/15/2017
  • Issued: 12/18/2018
  • Est. Priority Date: 08/20/2009
  • Status: Active Grant
First Claim
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1. A method of analyzing nucleic acids from a plurality of single cells, the method comprising:

  • (a) providing a sample comprising a plurality of single cells, wherein each single cell of the plurality of single cells comprises a plurality of sample polynucleotides;

    (b) generating a plurality of tagged polynucleotides from the plurality of sample polynucleotides, wherein each tagged polynucleotide comprises;

    (i) a sequence from a sample polynucleotide of the plurality of sample polynucleotides; and

    (ii) a multiplex identifier (MID) sequence comprising;

    I. a first tag sequence associated with the single cell from which the sample polynucleotide is derived, wherein the first tag sequence is a different sequence for different single cells in the plurality of single cells; and

    II. a second tag sequence distinguishing the sample polynucleotide from other sample polynucleotides derived from the same single cell;

    (c) sequencing the plurality of tagged polynucleotides to obtain a plurality of identified polynucleotide sequences;

    (d) using the first tag sequence to correlate the identified polynucleotide sequence with the single cell from which the identified polynucleotide sequence is derived; and

    (e) using the second tag sequence to correlate the identified polynucleotide sequence with the sample polynucleotide from which the identified polynucleotide sequence is derived.

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