Using random priming to obtain full-length V(D)J information for immune repertoire sequencing

US11773436B2 · kind B2 · utility

Assignee

• Becton, Dickinson and Company · US

Inventors

• Christina Chang · Menlo Park, US
• Margaret Nakamoto · Palo Alto, US

Key dates

• Filing date: Nov 6, 2020
• Grant date: Oct 3, 2023
• Expiry date: Jun 13, 2041

Classification

• Technology area (CPC C): Chemistry; Metallurgy
• CPC primary: C12Q2565/1025
• WIPO field: Biotechnology
• WIPO sector: Chemistry

Abstract

Disclosed herein include systems, methods, compositions, and kits for labeling nucleic acid targets in a sample. In some embodiments, nucleic acid targets (e.g., mRNAs) are initially barcoded on the 3′ end and are subsequently barcoded on the 5′ end following a template switching reaction and intermolecular and/or intramolecular hybridization and extension. 5′- and/or 3′-barcoded nucleic acid targets can serve as templates for amplification reactions and/or random priming and extension reactions to generate a sequencing library. The method can comprise generating a full-length sequence of the nucleic acid target by aligning a plurality of sequencing reads. Immune repertoire profiling methods are also provided in some embodiments.