title: Detection of mutations by fluorescence-assisted mismatch analysis (FAMA).
abstract: Fluorescence-assisted mismatch analysis (FAMA) methodology uses bifluorescent double-stranded DNA substrates to maximize the reliability and sensitivity of mutation-scanning procedures that rely on cleavage of mismatches using chemical. This unit presents a nested PCR procedure to fluorescently label each DNA strand, followed by chemical cleavage to detect the point mutations. Fluorescent end labeling with strand-specific fluorophores, electrophoretic separation of cleavage products in an automated Perkin-Elmer ABI 373 or 377 DNA sequencer and analysis using the Perkin-Elmer GENESCAN software expands sensitivity by highlighting weak signals through superimposition of strand-specific fluorescence electropherograms for different samples.
authors.affiliation: Institut Pasteur, Paris, France.
authors.forename: M
authors.initials: M
authors.lastname: Tosi
authors.forename: E
authors.initials: E
authors.lastname: Verpy
authors.forename: T
authors.initials: T
authors.lastname: Meo
mesh_terms: D020137:Base Pair Mismatch; D004252:DNA Mutational Analysis / Q000379:methods*; D005456:Fluorescent Dyes; D005826:Genetics, Medical; D006801:Humans; D009692:Nucleic Acid Heteroduplexes / Q000235:genetics / Q000302:isolation & purification; D016133:Polymerase Chain Reaction / Q000379:methods; D017422:Sequence Analysis, DNA
chemical_list: D005456:Fluorescent Dyes; D009692:Nucleic Acid Heteroduplexes