Amplification du polymorphisme rs10050860 (C<T) du gène ERAP1 par PCR en temps réel en utilisant la technologie TaqMan

Abstract

Real-time PCR, a technique that has revolutionized molecular biology by using fluorescent markers, offers faster detection, greater sensitivity and more accurate quantification of nucleic acids. In this work, we opted to use TaqMan probe technology, which uses specific probes to detect different genetic polymorphisms. We precisely quantified and amplified two DNA molecules extracted from two healthy individuals already available in a DNA bank at the LGMC Laboratory, USTO-MB by studying a single nucleotide polymorphism rs10050860 C<T of the ERAP1 gene at exon 12 of this gene. Quantification results showed that the concentrations of both DNAs were high, so dilution appeared to be necessary. Real-time PCR requires concentrations of no more than 50 ng/μL. The amplification results also enabled us to genotype the rs10050860 C<T polymorphism in the two healthy individuals. Secondly, real-time PCR enabled us to calculate Ct values for each sample. Finally, the use of real-time PCR with TaqMan technology proved to be a good, reliable and rapid approach, especially for the study of a genetic polymorphism such as the rs10050860 (C/T) of the ERAP1 gene explored in this work. Looking ahead, it would be interesting to study the rs10050860 polymorphism in patients, to better understand its role in autoimmune and inflammatory diseases.