TY - JOUR
T1 - Dual-color microchip electrophoresis with single-photon avalanche diodes
T2 - Application to mutation detection
AU - Stenirri, Stefania
AU - Cretich, Marina
AU - Rech, Ivan
AU - Restelli, Alessandro
AU - Ghioni, Massimo
AU - Cova, Sergio
AU - Ferrari, Maurizio
AU - Cremonesi, Laura
AU - Chiari, Marcella
PY - 2008
Y1 - 2008
N2 - 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.
AB - 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.
KW - Cystic fibrosis
KW - Microchip electrophoresis
KW - Mutation detection
KW - Single-photon avalanche diodes
UR - http://www.scopus.com/inward/record.url?scp=59349091683&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=59349091683&partnerID=8YFLogxK
U2 - 10.1002/elps.200800323
DO - 10.1002/elps.200800323
M3 - Article
C2 - 19130576
AN - SCOPUS:59349091683
VL - 29
SP - 4972
EP - 4975
JO - Electrophoresis
JF - Electrophoresis
SN - 0173-0835
IS - 24
ER -