Use of nucleic acid analogues in the inhibition of nucleic acid amplification

US5891625A · kind A · utility

Inventors

• Ole Buchardt · Frederikssund, DK
• Michael Egholm · Wayland, US
• Peter Nielsen · Kokkedal, DK
• Rolf Henrik Berg · Hørsholm, DK
• Christopher J. Stanley · London, GB

Key dates

• Filing date: Mar 10, 1995
• Grant date: Apr 6, 1999
• Expiry date: Mar 10, 2015

Classification

• Technology area (CPC C): Chemistry; Metallurgy
• CPC primary: C12Q1/686
• WIPO field: Biotechnology
• WIPO sector: Chemistry

Abstract

Nucleic acid analogues such as peptide-nucleic acids (PNAs) which hybridize strongly to nucleic acids are used to inhibit nucleic acid amplification procedures such as PCR. False positives in subsequent PCR assays are prevented by hybridizing a PNA to PCR amplification products. Assays capable of discriminating between single base mutants are conducted by using a PNA hybridizing to one of the two allelic forms to inhibit a PCR amplification of that form selectively. Asymmetric PCR amplifications are carried out by starting a PCR symmetrically using like quantities of forward and reverse primers, and, once the amplification is established, disabling one primer by hybridizing a PNA thereto.