Patent ID: 8080393

Claim:
A method for amplifying one or more oligonucleotides, said method comprising the steps of a) providing a nucleic acid probe (A) comprising one or more oligonucleotides and one or more double stranded nicking cassette(s) having complementary sequences, each nicking cassette comprising a double stranded nicking endonuclease recognition site having complementary nucleic acid sequences, wherein a nicking endonuclease capable of recognising said double stranded recognition site generates a nick in only one of the strands of the double stranded nucleic acid, b) circularising probe (A) as provided in step a) by self-templated hybridisation of the complementary nucleic acid sequences of the one or more nicking cassette(s) and ligating said circularised probe (A), c) providing a primer capable of recognising a target sequence in part of said circularised probe (A) obtained in step b), d) using the primer provided in step c) to effect rolling circle replication of said circularised probe (A), thereby providing a rolling circle product of probe (A) comprising one or more double stranded nicking cassette(s) having complementary sequences, each nicking cassette comprising a double stranded nicking endonuclease recognition site having complementary nucleic acid sequences, e) providing a nicking endonuclease capable of recognising double stranded nicking endonuclease recognition sites in the rolling circle product of probe (A), and nicking said double stranded nicking endonuclease recognition sites with the provided nicking endonuclease, thereby obtaining multiple copies of a probe (B) comprising a nucleic acid sequence complementary to the probe (A), f) circularising probe (B) as obtained in step e) by self-templated hybridisation of the complementary nucleic acid sequences of the one or more nicking cassette(s) and ligating said circularised probe (B) comprising a nucleic acid sequence complementary to the probe (A), g) providing a primer capable of recognising a target sequence in part of said circularised probe (B) obtained in step f), h) using the primer provided in step g) to effect rolling circle replication of said circularised probe (B), thereby providing a rolling circle product of probe (B) comprising one or more double stranded nicking cassette(s) having complementary sequences, each nicking cassette comprising a double stranded nicking endonuclease recognition site having complementary nucleic acid sequences, and i) providing a nicking endonuclease capable of recognising double stranded nicking endonuclease recognition sites in the rolling circle product of probe (B), and nicking said double stranded nicking endonuclease recognition sites with the provided nicking endonuclease, thereby obtaining multiple copies of a probe (A) comprising a nucleic acid sequence complementary to the probe (B).