Method for testing mutant gene through real-time polymerase chain reaction using DNA polymerase with inhibited activity of 5'-flap endonuclease

• Main Authors: LIM, SI KYU, CHA, SUN HO, KIM, JAE JONG
• Format: Patent
• Language: English
• Published: 21.07.2016
• Subjects: BEER; BIOCHEMISTRY; CHEMISTRY; COMPOSITIONS OR TEST PAPERS THEREFOR; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL ORENZYMOLOGICAL PROCESSES; ENZYMOLOGY; MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEICACIDS OR MICROORGANISMS; METALLURGY; MICROBIOLOGY; MUTATION OR GENETIC ENGINEERING; PROCESSES OF PREPARING SUCH COMPOSITIONS; SPIRITS; VINEGAR; WINE

The present invention relates to a method for detecting single nucleotide polymorphism (SNP) using a feature that the 5'-flap endonuclease (FEN) activity of DNA polymerase is inhibited when a probe complementarily binds to the end of a polymerase chain reaction (PCR) product. More specifically, the present invention relates to a novel method wherein it was verified that, when a probe used for a real-time PCR complementarily binds to the end site of a PCR product, the 5'-FEN activity of heat-resistant DNA polymerase for the probe is inhibited, and thus when such a feature is used to make a design such that an SNP site to be detected is located at the 5'-end site of the probe, the 5'-flap formation is induced according to the allele, thereby allowing effective SNP detection.