Nucleic acid amplification by a destabilization method

• Main Authors: LI YIMENG, MITRAN CATHERINE JENNIFER, GIBBS-DAVIS JULIANNE M, KAUSAR ABU, LAM JADE, MCKAY ROSALIE DAWN
• Format: Patent
• Language: English
• Published: 24.11.2015
• Subjects: BEER; BIOCHEMISTRY; CHEMISTRY; COMPOSITIONS OR TEST PAPERS THEREFOR; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL ORENZYMOLOGICAL PROCESSES; ENZYMOLOGY; FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIREDCHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERSFROM A RACEMIC MIXTURE; MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEICACIDS OR MICROORGANISMS; METALLURGY; MICROBIOLOGY; MUTATION OR GENETIC ENGINEERING; PROCESSES OF PREPARING SUCH COMPOSITIONS; SPIRITS; VINEGAR; WINE

The invention is directed to a method for isothermally amplifying a DNA sequence involving hybridizing a destabilizing DNA template to complementary nucleotide fragments to form a first nicked duplex; ligating the first nicked duplex to form a product duplex comprising the DNA sequence and the template, wherein the product duplex is capable of dissociating to release the DNA sequence and the template; and repeating these steps to generate multiple copies of the template and the DNA sequence. Further, the method may also involve hybridizing the DNA sequence to complementary destabilizing fragments or probes to form a second nicked duplex; ligating the second nicked duplex to form the product duplex comprising the DNA sequence and the template, wherein the product duplex dissociates to release the DNA sequence and the template; and repeating these steps to generate multiple copies of the template and the DNA sequence.