Patent ID: 11932849
Assignee: BECTON, DICKINSON AND COMPANY
Field: Biotechnology (Chemistry)
Classification: CPC C | IPC C

Claim 0:
1. A method of labeling nucleic acid targets in a sample, the method comprising:
contacting copies of a nucleic acid target with a plurality of oligonucleotide barcodes, wherein each oligonucleotide barcode comprises a first universal sequence, a molecular label, and a target-binding region capable of hybridizing to the copies of the nucleic acid target, wherein the nucleic acid target is an mRNA of a gene;
extending the plurality of oligonucleotide barcodes hybridized to the copies of the nucleic acid target to generate a plurality of first strand barcoded polynucleotides;
contacting random primers with the plurality of first strand barcoded polynucleotides, wherein each of the random primers comprises a second universal sequence, or a complement thereof,
extending the random primers hybridized to the plurality of first strand barcoded polynucleotides to generate a plurality of extension products;
amplifying the plurality of extension products using primers capable of hybridizing to the first universal sequence or complements thereof, and primers capable of hybridizing to the second universal sequence or complements thereof, thereby generating a first plurality of barcoded amplicons; and
synthesizing a plurality of gene-specific barcoded amplicons using the plurality of first strand barcoded polynucleotides as templates, wherein synthesizing a plurality of gene-specific barcoded amplicons comprises PCR amplification using primers capable of hybridizing to the first universal sequence, or a complement thereof, and a gene-specific primer.