METHODS FOR ANALYZING NUCLEIC ACIDS FROM SINGLE CELLS
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; and
II. a second tag sequence distinguishing the sample polynucleotide from other pluralities of 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 polynucleotide sequence is derived; and
(e) using the second tag sequence to correlate the identified polynucleotide sequence with the plurality of sample polynucleotides from which the polynucleotide sequence is derived.
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Abstract
Aspects of the present invention include analyzing nucleic acids from single cells using methods that include using tagged polynucleotides containing multiplex identifier sequences.
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Citations
6 Claims
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1. A method of analyzing nucleic acids from a plurality of single cells, the method comprising:
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(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; and II. a second tag sequence distinguishing the sample polynucleotide from other pluralities of 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 polynucleotide sequence is derived; and (e) using the second tag sequence to correlate the identified polynucleotide sequence with the plurality of sample polynucleotides from which the polynucleotide sequence is derived. - View Dependent Claims (2, 3, 4, 5, 6)
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Specification