Patent ID: 9657339
Date: 2017-05-23
CPC Classifications: C12Q

Claim:
1. A method for amplifying at least one target nucleic acid in a nucleic acid sample, comprising: (a) fragmenting the nucleic acid sample to create at least one target fragment comprising said target nucleic acid and comprising two probe-complementary portions, wherein at least one of said two probe-complementary portions is located at an end of the target fragment; (b) contacting said fragmented nucleic acid sample with at least one probe which is provided with an immobilization moiety and optionally is immobilized on a solid phase by means of said moiety, and which comprises two target fragment-complementary portions which are complementary in sequence to the probe-complementary portions of the target fragment, wherein said portions of the probe may be adjacent, or separated by an intervening non-target fragment-complementary portion; (c) rendering the fragmented nucleic acid sample single-stranded, wherein the step may occur before, contemporaneously with or after step (b); (d) allowing the probe-complementary portions of the target fragment to hybridize with the target-fragment complementary portions of the probe; (e) if the probe provided in step (b) is not immobilized, immobilizing the probe-target fragment hybrid on a solid phase by means of said immobilization moiety; (f) separating non-immobilized nucleic acid fragments from the solid phase; (g) contacting the solid phase with a ligase to ligate, directly or indirectly, ligatable 5′ and 3′ ends of the target fragment whereby the target fragment is circularized, wherein where one of said probe-complementary portions of the target fragment is not located at an end of the target fragment, the ligatable end, other than that at which a probe-complementary portion is located, is created by the action of a flap endonuclease or an exonuclease; and (h) amplifying said circularized target fragment.