Method for amplification of target nucleic acid

US9657339B2 · kind B2 · utility

Assignee

• AGILENT TECHNOLOGIES, INC. · US

Inventors

• Fredrik Roos · Stuvsta-Snättringe, SE
• Henrik Johansson · Uppsala, SE
• Magnus Isaksson · Täby, SE
• Mats Nilsson · Uppsala, SE
• Olle (Olof) Ericsson · Uppsala, SE
• Simon Fredriksson · Stockholm, SE

Key dates

• Filing date: Dec 3, 2010
• Grant date: May 23, 2017
• Expiry date: Apr 23, 2033

Classification

• Technology area (CPC C): Chemistry; Metallurgy
• CPC primary: C12Q1/6837
• WIPO field: Biotechnology
• WIPO sector: Chemistry

Abstract

A method for amplifying a target nucleic acid is disclosed, which includes: (a) fragmenting a nucleic acid sample to create a target fragment comprising a target nucleic acid and two probe-complementary portions; (b) contacting said fragmented nucleic acid sample with a probe comprising two target fragment-complementary portions complementary to the probe-complementary portions of the target fragment; (c) rendering the fragmented nucleic acid sample single-stranded; (d) allowing the probe-complementary portions to hybridise with the target-fragment complementary portions; (e) if the probe in step (b) is not immobilised, immobilising the probe-target fragment hybrid on a solid phase via immobilisation moiety; (f) separating non-immobilised nucleic acid fragments from the solid phase; (g) contacting the solid phase with a ligase to ligate ligatable 5′ and 3′ ends of the target fragment whereby the target fragment is circularized; and (h) amplifying said circularized target fragment.