title: A combined allele-specific PCR and RFLP assay to detect the 35delG mutation in the Connexin 26 gene.
abstract: Mutations in the Connexin-26 (specified GJB2) gene have been shown to be a major cause of nonsyndromic recessive deafness (NSRD), and a single mutation 35delG in the GJB2 gene accounts for the majority of cases of NSRD. For diagnostic analyses and for scientific studies of large numbers of patients, fast and economic assays that can be performed with standard polymerase chain reaction (PCR) instruments are highly desirable. We have developed an allele-specific amplification (ASA)-based restriction fragment length polymorphism (RFLP) assay. We evaluated the multiplex method for its ability to 35delG mutation. Our method is a stable, reproducible and concordend with previously reported PCR-RFLP assays.
authors.affiliation: Department of Molecular Biology and Genetics, Bogazici University, 34342, Istanbul, Turkey. ibrahim.baris@boun.edu.tr
authors.forename: Ibrahim
authors.initials: I
authors.lastname: Baris
authors.forename: Vedat
authors.initials: V
authors.lastname: Koksal
authors.forename: Ozdal
authors.initials: O
authors.lastname: Etlik
mesh_terms: D001483:Base Sequence; D000072259:Connexin 26; D017630:Connexins / Q000235:genetics*; D004252:DNA Mutational Analysis; D017931:DNA Primers; D003638:Deafness / Q000235:genetics*; D005820:Genetic Testing; D006801:Humans; D008969:Molecular Sequence Data; D021141:Nucleic Acid Amplification Techniques; D017354:Point Mutation; D016133:Polymerase Chain Reaction / Q000379:methods*; D012150:Polymorphism, Restriction Fragment Length; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity
chemical_list: D017630:Connexins; D017931:DNA Primers; C000607042:GJB2 protein, human; D000072259:Connexin 26