title: Dual-color microchip electrophoresis with single-photon avalanche diodes: application to mutation detection.
abstract: A novel microchip electrophoresis instrument based on single-photon avalanche diodes was used for the molecular characterization of mutations in disease genes. The identification of the main mutation causing cystic fibrosis, named DeltaF508, by the Amplification Refractory Mutation System was used to validate the technology. In our implemented protocol the wild-type and mutant allele-specific primers are labeled with Cy5 and Cy5.5, respectively. The protocol enables the amplification of the DNA sample in a single PCR. The genotype was deduced from the fluorescence of the amplicons run in the CE microchip. Validation on 15 DNA samples from either homozygous wild-type or heterozygous and homozygous mutated control subjects proved the complete reliability of the system, thus confirming its high diagnostic potential.
authors.affiliation: Genomic Unit for the Diagnosis of Human Pathologies, San Raffaele Scientific Institute, Milan, Italy.
authors.forename: Stefania
authors.initials: S
authors.lastname: Stenirri
authors.forename: Marina
authors.initials: M
authors.lastname: Cretich
authors.forename: Ivan
authors.initials: I
authors.lastname: Rech
authors.forename: Alessandro
authors.initials: A
authors.lastname: Restelli
authors.forename: Massimo
authors.initials: M
authors.lastname: Ghioni
authors.forename: Sergio
authors.initials: S
authors.lastname: Cova
authors.forename: Maurizio
authors.initials: M
authors.lastname: Ferrari
authors.forename: Laura
authors.initials: L
authors.lastname: Cremonesi
authors.forename: Marcella
authors.initials: M
authors.lastname: Chiari
mesh_terms: D004247:DNA / Q000032:analysis / Q000235:genetics; D004252:DNA Mutational Analysis / Q000379:methods*; D046209:Electrophoresis, Microchip / Q000295:instrumentation* / Q000379:methods; D009154:Mutation / Q000235:genetics*; D017785:Photons
chemical_list: D004247:DNA