Methods and compositions for tagging and analyzing samples
First Claim
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1. A method of tagging target oligonucleotides comprising:
- performing an in vitro transcription reaction on DNA molecules partitioned within a plurality of separate compartments, wherein each DNA molecule comprises an RNA polymerase promoter region, a randomer tag region, and a hybridization region capable of hybridizing to one or more target oligonucleotides, thereby obtaining compartments comprising RNA comprising the RNA polymerase promoter region, the randomer tag region and the hybridization region;
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;
hybridizing the RNA to the target oligonucleotides; and
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
28 Claims
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1. A method of tagging target oligonucleotides comprising:
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performing an in vitro transcription reaction on DNA molecules partitioned within a plurality of separate compartments, wherein each DNA molecule comprises an RNA polymerase promoter region, a randomer tag region, and a hybridization region capable of hybridizing to one or more target oligonucleotides, thereby obtaining compartments comprising RNA comprising the RNA polymerase promoter region, the randomer tag region and the hybridization region; 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; hybridizing the RNA to the target oligonucleotides; and 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, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28)
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7. The method of wherein the target oligonucleotides are obtained by lysing one or more cells within individual compartments of the set of compartments to liberate cellular oligonucleotides, wherein the cellular oligonucleotides are the target oligonucleotides.
Specification