Compositions and methods for molecular labeling
First Claim
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1. A method of making a barcode library comprising:
- obtaining a first nucleic acid construct wherein the first construct comprises a forward sequence specific primer, a first barcode sequence, and a first universal primer recognition site;
compartmentalizing the first construct into a fluid compartment;
introducing a second nucleic acid construct into the fluid compartment, wherein the second construct comprises a reverse sequence specific primer, a second barcode sequence, and a second universal primer recognition site;
introducing a third construct into the fluid compartment and hybridizing the third construct to the first construct to form a hybrid construct; and
ligating the hybrid construct to form a new construct.
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Abstract
The invention provides barcode libraries and methods of making and using them including obtaining a plurality of nucleic acid constructs in which each construct comprises a unique N-mer and a functional N-mer and segregating the constructs into a fluid compartments such that each compartment contains one or more copies of a unique construct. The invention further provides methods for digital PCR and for use of barcode libraries in digital PCR.
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Citations
14 Claims
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1. A method of making a barcode library comprising:
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obtaining a first nucleic acid construct wherein the first construct comprises a forward sequence specific primer, a first barcode sequence, and a first universal primer recognition site; compartmentalizing the first construct into a fluid compartment; introducing a second nucleic acid construct into the fluid compartment, wherein the second construct comprises a reverse sequence specific primer, a second barcode sequence, and a second universal primer recognition site; introducing a third construct into the fluid compartment and hybridizing the third construct to the first construct to form a hybrid construct; and ligating the hybrid construct to form a new construct. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14)
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Specification