Processes and systems for preparation of nucleic acid sequencing libraries and libraries prepared using same

US10557158B2 · kind B2 · utility

Assignee

• 10x Genomics, Inc. · US

Inventors

• Paul Hardenbol · San Francisco, US
• Pranav Patel · Palo Alto, US
• Benjamin Hindson · Pleasanton, US
• Paul William Wyatt · Pleasanton, US
• Keith Bjornson · Fremont, US
• Indira Wu · Redwood City, US
• Kamila Belhocine · Palo Alto, US

Key dates

• Filing date: Dec 20, 2018
• Grant date: Feb 11, 2020
• Expiry date: Dec 20, 2038

Classification

• Technology area (CPC B): Performing Operations; Transporting
• CPC primary: B01J2219/00722
• WIPO field: Biotechnology
• WIPO sector: Chemistry

Abstract

This disclosure provides methods for preparing a sequencing library including the steps of providing a template nucleic acid sequence, dNTPs, dUTP, a primer, a polymerase, a dUTP excising enzyme, and a plurality of beads including oligonucleotide adapter sequence segments; amplifying the template nucleic acid with the polymerase, dNTPs, dUTP and random hexamer to provide a complementary nucleic acid sequence including occasional dUTPs; and excising the incorporated dUTPs with the dUTP excising enzyme to provide nicks in the complementary nucleic acid sequence to provide a sequencing library.