Materials and methods for detection of nucleic acids

US7422850B2 · kind B2 · utility

Assignee

• Eragen Biosciences, Inc. · US

Inventors

• David J. Marshall · Elmore, AU
• James R. Prudent · Madison, US
• Christopher B. Sherrill · Madison, US
• Gideon Shapiro · Gainesville, US
• Jennifer K. Grenier · Madison, US
• Craig S. Richmond · Madison, US
• Simona Jurczyk · Gainesville, US
• Jerod PTACIN · Beloit, US

Key dates

• Filing date: May 18, 2001
• Grant date: Sep 9, 2008
• Expiry date: Mar 9, 2022

Classification

• Technology area (CPC C): Chemistry; Metallurgy
• CPC primary: C12Q2600/156
• WIPO field: Biotechnology
• WIPO sector: Chemistry

Abstract

Assays using non-natural bases are described. In one embodiment, the method involves contacting a sample suspected of containing the target nucleic acid with a polymerase and first and second primers; amplifying the target nucleic acid, if present in the sample, by PCR using the first and second primers to generate an amplification product having a double-stranded region and a single-stranded region that comprises the non-natural base; contacting the sample with a reporter comprising a label and a non-natural base that is complementary to the non-natural base of the single-stranded region; annealing at least a portion of the reporter to the single-stranded region of the amplification product; cleaving, after annealing, at least a portion of the reporter to release at least one reporter fragment; and correlating the release of the at least one reporter fragment with the presence of the target nucleic acid in the sample. The invention also provides corresponding kits for use in detecting target nucleic acids in a sample. Alternatively, the reporter can be incorporated into the amplification product rather than annealing and then cleaving.