Safe sequencing system

US11180803B2 · kind B2 · utility

Assignee

• THE JOHNS HOPKINS UNIVERSITY · US

Inventors

• Bert Vogelstein · Baltimore, US
• Kenneth W. Kinzler · Baltimore, US
• Nickolas Papadopoulos · Towson, US
• Isaac A. Kinde · Beaumont, US

Key dates

• Filing date: Aug 18, 2016
• Grant date: Nov 23, 2021
• Expiry date: Aug 18, 2036

Classification

• Technology area (CPC C): Chemistry; Metallurgy
• CPC primary: C12Q2600/158
• WIPO field: Biotechnology
• WIPO sector: Chemistry

Abstract

Though massively parallel sequencing instruments are in principle well-suited to the task of identification of mutations present in a small fraction of DNA templates, the error rates in such instruments are generally too high to allow confident identification of rare variants. An approach that can substantially increase the sensitivity of massively parallel sequencing instruments for this purpose is called “Safe-SeqS” for (Safe-Sequencing System) and includes (i) assignment of a unique identifier (UID) to each template molecule; (ii) amplification of each uniquely tagged template molecule to create UID-families; and (iii) redundant sequencing of the amplification products. PCR fragments with the same UID are truly mutant (“super-mutants”) if ≥95% of them contain the identical mutation.