Compositions and methods for molecular labeling
AssigneeBio-Rad Laboratories, Inc.
InventorsMichael L. Samuels, Jeffrey Charles Olson, Andrew Watson, Keith Brown, Darren R. Link
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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.
Claims13
A method for analyte detection, the method comprising the steps of:
- exposing target molecules of cells from a biological sample to a plurality of antibodies comprising antibodies that are linked to a target-specific oligonucleotide, thereby forming a plurality of complexes that comprise a target molecule bound to an antibody linked to a target-specific oligonucleotide, wherein the target-specific oligonucleotide linked to an antibody in a given complex of the plurality of complexes comprises a nucleic acid that includes a target-specific sequence that identifies the target molecule bound by the antibody of that complex;
- isolating an individual cell comprising one or more complexes of the plurality of complexes in a compartment;
- attaching a compartment-specific barcode to a target-specific oligonucleotide of one or more of the complexes within the compartment;
- and
- obtaining sequence information of the compartment-specific barcode and the target-specific sequence of the one or more complexes to identify one or more of the target molecules and assign the one or more target molecules to the individual cell.
A method for identifying a target molecule, comprising the steps of:
- exposing a cell comprising a target molecule to an antibody that binds the target molecule and comprises an oligonucleotide comprising a sequence that identifies the antibody;
- isolating the cell in an aqueous partition with a partition barcode;
- attaching the partition barcode to the oligonucleotide from the antibody,
- forming a nucleic acid that includes the sequence that identifies the antibody and the partition barcode;
- obtaining sequence information of the nucleic acid that includes the sequence that identifies the antibody and the partition barcode;
- and
- based on the sequence information of the sequence that identifies the antibody, determining the identity of the target molecule, and
- based on the sequence information of the partition barcode, assigning the target molecule to the cell.
Dependencies are inferred from the claim text.