System and method for determining copies-per-unit-volume using PCR and flow control of droplets

US10557174B2 · kind B2 · utility

Assignee

• Life Technologies Corporation · US

Inventors

• Gordon A. Janaway · Hayward, US
• Mark Andersen · San Mateo, US
• Kornelija Zgonc · Carlsbad, US
• Michael C. Pallas · San Bruno, US
• Marcin Sikora · Foster City, US
• Casey R. McFarland · San Francisco, US
• Ferrier N. Le · San Jose, US
• Haopeng Wang · Ellicott City, US
• Jian Gong · Shanghai, CN
• Gothami Padmabandu · San Diego, US

Key dates

• Filing date: Mar 25, 2016
• Grant date: Feb 11, 2020
• Expiry date: Jul 3, 2037

Classification

• Technology area (CPC C): Chemistry; Metallurgy
• CPC primary: C12Q2600/178
• WIPO field: Measurement
• WIPO sector: Instruments

Abstract

Methods and systems for quantification of a target nucleic acid in a sample are provided. The method includes forming a plurality of discrete sample portions. Each of the plurality of discrete sample portions comprising a portion of the sample, and a reaction mixture. The method further includes amplifying the plurality of discrete sample portions to form a plurality of discrete processed sample portions. At least one discrete processed sample portion containing nucleic acid amplification reaction products. Fluorescence signals are detected from the at least one of the plurality of discrete processed sample portions to determine a presence of the at least one target nucleic acid. The method also includes determining the respective volumes of the plurality of the plurality of discrete processed sample portions, and estimating the number of copies-per-unit-volume of the at least one target nucleic acid in the sample. Estimating the number of copies-per-unit-volume is based on the number of discrete processed sample portions determined to contain the at least one target nucleic acid therein.