title: Simultaneous genotyping of multiple somatic mutations by using a clamping PNA and PNA detection probes.
abstract: It has been very difficult to detect and quantify multiple somatic mutations simultaneously in single-tube qPCR. Here, a novel method for simultaneous detection of multiple mutations and a melting curve analysis was developed by using clamping PNA and detection PNA probes. Each PNA probe was designed to have a specific melting temperature by the introduction of γ-PNA monomer. This technique was successfully applied to the detection of six genotypes in two different mutations of K-RAS at the same time. Such simultaneous analysis of an amplified curve and a melting curve in qPCR can be widely used for the early diagnosis of cancer and determining the prognosis of drug treatments.
authors.affiliation: Research Institute, Panagene Inc. 54 Techno10-ro, Yuseong-gu, Daejeon, 305-510 (Republic of Korea).
authors.forename: Yong-Tae
authors.initials: YT
authors.lastname: Kim
authors.forename: Jin Woo
authors.initials: JW
authors.lastname: Kim
authors.forename: Sung Kee
authors.initials: SK
authors.lastname: Kim
authors.forename: Goon Ho
authors.initials: GH
authors.lastname: Joe
authors.forename: In Seok
authors.initials: IS
authors.lastname: Hong
mesh_terms: D000409:Alanine / Q000737:chemistry; D002460:Cell Line; D019281:Dimerization; D011905:Genes, ras / Q000235:genetics*; D060005:Genotyping Techniques / Q000379:methods; D006367:HeLa Cells; D006801:Humans; D008239:Lysine / Q000737:chemistry; D015335:Molecular Probes; D009154:Mutation; D020135:Peptide Nucleic Acids / Q000235:genetics*; D016133:Polymerase Chain Reaction / Q000379:methods*; D013696:Temperature
chemical_list: D015335:Molecular Probes; D020135:Peptide Nucleic Acids; D008239:Lysine; D000409:Alanine
keywords: cancer; gamma-PNA; multiple genotyping; peptide nucleic acid; quantitative PCR; somatic mutations