Method and system for ultra-high dynamic range nucleic acid quantification

US10081017B2 · kind B2 · utility

Assignee

• The Regents of the University of California · US

Inventors

• Abraham P. Lee · Irvine, US
• Andrew C. Hatch · Irvine, US

Key dates

• Filing date: Oct 8, 2015
• Grant date: Sep 25, 2018
• Expiry date: May 16, 2036

Classification

• Technology area (CPC B): Performing Operations; Transporting
• CPC primary: B01L2300/0864
• WIPO field: Chemical engineering
• WIPO sector: Chemistry

Abstract

A method of quantifying nucleic acids in a sample includes generating a plurality of droplets in oil within a microfluidic device, wherein at least some of the droplets comprise a nucleic acid, amplification reagents, and a fluorescent probe or dyes contained therein. The droplets are delivered to a collection chamber to form an array of droplets. The droplets are subject to thermal cycling within the collection chamber a plurality of times to perform nucleic acid amplification within the droplets. The array of droplets is imaged during the plurality of thermal cycles as well as at a thermal cycle endpoint. An initial concentration of nucleic acid in the sample is calculated based on at least one of: a ratio of aqueous phase droplets exhibiting fluorescence within the array at the thermal cycle endpoint or a cycle threshold (Ct) of one or more aqueous phase droplets within the array.