Method for the efficiency-corrected real-time quantification of nucleic acids

US6691041B2 · kind B2 · utility

Assignee

• Roche Molecular Systems, Inc. · US

Inventors

• Gregor Sagner · Penzberg, DE
• Karim Tabiti · Pocking, DE
• Martin Gutekunst · Eberfing, DE
• Richie Soong · Tutzing, DE

Key dates

• Filing date: Mar 30, 2001
• Grant date: Feb 10, 2004
• Expiry date: Mar 30, 2021

Classification

• Technology area (CPC C): Chemistry; Metallurgy
• CPC primary: C12Q1/6851
• WIPO field: Biotechnology
• WIPO sector: Chemistry

Abstract

The present invention concerns a method for the quantification of a target nucleic acid in a sample comprising the following steps: (i) determination of the amplification efficiency of the target nucleic acid under defined amplification conditions, (ii) amplification of the target nucleic acid contained in the sample under the same defined reaction conditions, (iii) measuring the amplification in real-time, (iv) quantification of the original amount of target nucleic acid in the sample by correction of the original amount derived from step (iii) with the aid of the determined amplification efficiency. The efficiency correction of PCR reactions according to the invention for the quantification of nucleic acids can be used for absolute quantification with the aid of an external or internal standard as well as for relative quantification compared to the expression of housekeeping genes.