The Development and Optimization of Primer Sets Used to Study the Relative Expression of Androgen Receptor Gene in Turkey (Meleagris gallopavo)

Abstract

The Androgen Receptor (AR) Gene’s expression is essential during puberty and testes maturation, which is also used as a reference for Turkey’s breeding program. Therefore, this study aims to develop and optimize two primer sets for studying relative expression using qPCR technology. The primers were designed to amplify specific regions in the AR gene as the main target, and the β-actin gene as an internal control. They were tested using in-silico and amplicon sequencing, as well as efficiency calculation with a constructed standard curve from serially diluted reactions. Based on the sequencing methods, the primers amplified the corresponding regions of the respective targets. The primer for AR gene had an efficiency of 98.03%, a slope of -3.37, and an R2 of 0.995, while that of the β-actin gene had an efficiency of 98.01%, a slope of -3.371, and an R2 of 0.999. The two efficiencies exceeded the standard (93 -103%) value and the melting curve analysis showed that no non-specific amplification was discovered for both primers. According to the tests, the primers are suggested as acceptable to be used for the relative expression study of the AR gene in Turkey.