Methods and compositions for tagging and analyzing samples
DCFirst 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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Abstract
The invention relates to methods of tagging analytes in a sample.
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Citations
25 Claims
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1. A method of tagging target oligonucleotides comprising:
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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. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25)
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Specification