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Digital counting of individual molecules by stochastic attachment of diverse labels

  • US 10,392,661 B2
  • Filed: 12/13/2018
  • Issued: 08/27/2019
  • Est. Priority Date: 12/15/2009
  • Status: Active Grant
First Claim
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1. A method, comprising:

  • (a) attaching to one or more occurrences of a first target molecule and a second target molecule in a sample (i) a first label sequence randomly selected from a first set of label sequences and (ii) a second label sequence randomly selected from a second set of label sequences, thereby generating for each occurrence of the first target molecule and the second target molecule a new first molecule and a new second molecule, respectively,wherein each new first molecule and new second molecule comprises a copy of the first target molecule and a copy of the second target molecule, respectively, and a combination label sequence comprising a first label sequence and a second label sequence,wherein n1 and n2 are the numbers of the one or more occurrences of the first target molecule and the second target molecule, respectively, in the sample,wherein combinations of each of the first set of label sequences and each of the second set of label sequences comprises m different combination label sequences, andwherein the ratio of the greater of n1 and n2 to m is smaller than 0.2; and

    (b) detecting the new first molecules and the new second molecules by detecting the combination label sequences present on the new first molecules and the new second molecules, wherein the detected new first molecules and the detected new second molecules indicate the number of the one or more occurrences of the first target molecule and the number of the one or more occurrences of the second target molecule, respectively.

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