A PCR-RFLP test for simultaneous detection of two single-nucleotide insertions in the Connexin-26 gene promoter.

Comparisons of Connexin-26 (GJB2) gene sequences available in the GenBank data base indicate the presence of a polymorphism in the promoter, but no easy method is available for the detection of this polymorphism. We have developed a PCR-RFLP test for simultaneous detection of two single nucleotide insertions (G and A) in the GJB2 promoter. The test is based on amplification of a 146-bp DNA fragment, which was digested with Mae I to detect the G insertion in the promoter.

Absence of deafness-associated connexin-26 (GJB2) gene mutations in the Omani population.

We have investigated the prevalence of mutations in the connexin 26 (GJB2) gene in Omani population using both PCR-RFLP and direct DNA sequencing methods. Two common GJB2 gene mutations (35delG and 167delT) were screened in 280 healthy controls and 95 deaf patients using two different PCR-RFLP methods. To investigate other GJB2 mutations, we have amplified and sequenced DNA from 51 unrelated deaf patients and 17 control subjects.

A polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) test to detect the common mutation (35delG) in the connexin-26 gene.

Objective: To develop a polymerase chain reaction (PCR) based test for the detection of a common frame-shift mutation (35delG) in the connexin-26 (GJB2) gene, and to investigate the status of this mutation in Oman.

Method: A PCR test, based on site-directed mutagenesis, was developed for the 35delG mutation. A mutagenesis primer generated an EcoN I site in a short (87 bp) DNA fragment amplified from the connexin-26 gene.

A seminested PCR test for simultaneous detection of two common mutations (35delG and 167delT) in the connexin-26 gene.

Background: Several mutations described in the connexin-26 gene cause nonsyndromic autosomal recessive deafness (NARD). The prevalence of two frame-shift mutations, known as 35delG and 167delT, was relatively high in patients with NARD from different populations.

Methods And Results: A seminested PCR test has been developed for simultaneous detection of two common mutations in the connexin-26 gene.