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Methods and compositions for tagging and analyzing samples

DC
  • US 10,058,839 B2
  • Filed: 03/14/2014
  • Issued: 08/28/2018
  • Est. Priority Date: 03/15/2013
  • Status: Active Grant
First Claim
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1. A method of tagging target oligonucleotides comprising:

  • a. partitioning DNA into a plurality of compartments, wherein the DNA comprises an RNA polymerase promoter sequence, a tag sequence and a hybridization region capable of hybridizing to one or more target oligonucleotides, and wherein the plurality of compartments or a subset thereof further comprises an RNA polymerase and ribonucleotide triphosphates;

    b. performing an in vitro transcription reaction on the DNA within the compartments, thereby obtaining compartments comprising RNA, wherein the RNA comprises the tag sequence and the hybridization region;

    c. merging the interior of the compartments comprising RNA with the interior of a set of compartments comprising target oligonucleotides, wherein prior to the merging, the compartments comprising target oligonucleotides are physically separate from the compartments comprising RNA;

    d. hybridizing the RNA to the target oligonucleotides, wherein the hybridization region of the RNA hybridizes to a region of the target oligonucleotides; and

    e. performing a reaction to attach a sequence corresponding to the RNA to the target oligonucleotides, thereby tagging the target oligonucleotides.

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