×

System and Methods for Massively Parallel Analysis of Nucleic Acids in Single Cells

  • US 20140057799A1
  • Filed: 12/16/2011
  • Published: 02/27/2014
  • Est. Priority Date: 12/16/2010
  • Status: Abandoned Application
First Claim
Patent Images

1. A method for analyzing at least two nucleic acid sequences in a single cell contained within a population of at least 10,000 cells, comprising:

  • providing a first set of nucleic acid probes, the first set comprising a first probe comprising a sequence that is complementary to a first target nucleic acid subsequence, a second probe comprising a sequence that is complementary to a second subsequence of the first target nucleic acid and a second sequence that is complementary to an exogenous sequence, a third probe comprising the exogenous sequence and a sequence that is complementary to a first subsequence of a second target nucleic acid, and a fourth probe comprising a sequence that is complementary to a second subsequence of the second target nucleic acid sequence, wherein the first target nucleic acid or the second target nucleic acid comprises an endogenous sequence;

    isolating the single cells with at least one set of nucleic acid probes;

    amplifying the first and second target nucleic acid sequences independently, wherein the first target nucleic acid sequence is amplified using the first probe and the second probe, and wherein the second target nucleic acid sequence is amplified using the third probe and the fourth probe;

    hybridizing the exogenous sequence to its complement;

    amplifying the first target nucleic acid sequence, the second target nucleic acid sequence, and the exogenous sequence using the first and fourth probes, thereby generating a fused complex; and

    performing a bulk sequencing reaction to generate sequence information for at least 100,000 fused complexes from at least 10,000 cells within the population of cells, wherein the sequence information is sufficient to co-localize the first target nucleic acid sequence and the second target nucleic acid sequence to a single cell from the population of at least 10,000 cells.

View all claims
  • 3 Assignments
Timeline View
Assignment View
    ×
    ×