Dual-color microchip electrophoresis with single-photon avalanche diodes: Application to mutation detection

Stefania Stenirri, Marina Cretich, Ivan Rech, Alessandro Restelli, Massimo Ghioni, Sergio Cova, Maurizio Ferrari, Laura Cremonesi, Marcella Chiari

Research output: Contribution to journalArticle

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 ΔF508, 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.

Original languageEnglish
Pages (from-to)4972-4975
Number of pages4
JournalElectrophoresis
Volume29
Issue number24
DOIs
Publication statusPublished - 2008

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Keywords

  • Cystic fibrosis
  • Microchip electrophoresis
  • Mutation detection
  • Single-photon avalanche diodes

ASJC Scopus subject areas

  • Biochemistry
  • Clinical Biochemistry

Cite this

Stenirri, S., Cretich, M., Rech, I., Restelli, A., Ghioni, M., Cova, S., Ferrari, M., Cremonesi, L., & Chiari, M. (2008). Dual-color microchip electrophoresis with single-photon avalanche diodes: Application to mutation detection. Electrophoresis, 29(24), 4972-4975. https://doi.org/10.1002/elps.200800323