Digital amplification

US6753147B2 · kind B2 · utility

Assignee

• THE JOHNS HOPKINS UNIVERSITY · US

Inventors

• Bert Vogelstein · Baltimore, US
• Kenneth W. Kinzler · Baltimore, US

Key dates

• Filing date: Oct 12, 2001
• Grant date: Jun 22, 2004
• Expiry date: Oct 12, 2021

Classification

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

Abstract

The identification of pre-defined mutations expected to be present in a minor fraction of a cell population is important for a variety of basic research and clinical applications. The exponential, analog nature of the polymerase chain reaction is transformed into a linear, digital signal suitable for this purpose. Single molecules can be isolated by dilution and individually amplified; each product is then separately analyzed for the presence of pre-defined mutations. The process provides a reliable and quantitative measure of the proportion of variant sequences within a DNA sample.