Establishment and rapid detection of a heterozygous missense mutation in the CACNA1F gene by ARMS technique with double-base mismatched primers

Abstract

ABSTRACT. Retinitis pigmentosa (RP) is a retinal degenerative disorder that often causes complete blindness. Mutations of more than 50 genes have been identified as associated with RP, including the CACNA1F gene. In a recent study, by employing next-generation sequencing, we identified a novel mutation in the CACNA1F gene. In this study, we used the amplification refractory mutation system (ARMS) and identified a single nucleotide change c.1555C>T in exon 13 of the CACNA1F gene, leading to the substitution of arginine by tryptophan (p.R519W) in a Chinese individual affected by RP. Establishment and rapid detection of CACNA1F gene by ARMS This study actually confirms this novel mutation, and establishes the ARMS technique for the detection of mutations in RP.

Evaluation of amplification refractory mutation system (ARMS) technique for quick and accurate prenatal gene diagnosis of <em>CHM</em> variant in choroideremia

“…Previously, Fu et al and Moghadam et al successfully used ARMS to diagnose β-thalassemia prenatally in China and Iran, respectively; 18 , 31 Chiu et al applied this method to quantify given mutant mtDNA heteroplasmies, 32 whereas Newton et al used ARMS for antenatal diagnosis of cystic fibrosis. 33 Recently, Yang et al performed rapid gene diagnosis for retinitis pigmentosa, 34 and Singh et al performed prenatal gene diagnosis for sickle cell disease by ARMS. 35 More recently, Aquino et al performed ARMS-PCR to detect the most common mutations of the CFTR gene in Peruvian patients with cystic fibrosis.…”

Identification of the origin of marker chromosomes by two-color fluorescence in situ hybridization and polymerase chain reaction in azoospermic patients

“…The cell pellets were resuspended in 3 mL 1X nucleic lysis buffer (1 mM Tris, pH 8.2, 40 mM NaCl, 0.2 mM EDTA) and vortexed. Sodium dodecyl sulfate (300 μL, 10%) and proteinase K (10 μL, 20 mg/mL) were added, mixed, incubated at 55°C for 3-5 h or 37°C overnight, and finally the DNAs were routinely extracted by phenol/chloroform (Fu et al, 2002;Fu, 2012;Long et al, 2014;Khan et al, 2015;Yang et al, 2015).…”