Detection of the DeltaF508 mutation in the CFTR gene by means of time-resolved fluorescence methods.
A rapid recognition in the base sequence of nucleic acids is an important prerequisite toward the diagnosis of genetic diseases and their carrier states. We have developed a hybridisation method in which a fluorescently labeled oligonucleotide is used to detect point mutations in a target by a simple fluorescence lifetime analysis of the emission of the fluorescent label. We applied this method to detect the deltaF508 mutation in the cystic fibrosis transmembrane conductance regulator (CFTR) gene in a model system and with biologically derived PCR product and discuss the potential generality of this method.
Kirschstein
S
S
Institute of Molecular Biotechnology, Department of Single Cell and Single Molecule Techniques, Jena, Germany. omar@imb-jena.de
Winter
S
S
Turner
D
D
Löber
G
G
D001483:Base Sequence; D001896:Boron Compounds; D019005:Cystic Fibrosis Transmembrane Conductance Regulator / Q000235:genetics*; D017931:DNA Primers; D005456:Fluorescent Dyes; D017404:In Situ Hybridization, Fluorescence; D017354:Point Mutation; D016133:Polymerase Chain Reaction; D013050:Spectrometry, Fluorescence
C095489:4,4-difluoro-4-bora-3a,4a-diaza-s-indacene; D001896:Boron Compounds; D017931:DNA Primers; D005456:Fluorescent Dyes; D019005:Cystic Fibrosis Transmembrane Conductance Regulator