Primer characterization of in-house real time PCR (RT-PCR) for BCL2 gene using saliva sample

Abstract

In organisms, cells perform apoptosis to remove damaged or mutated cells from the body. The Bcl-2 family protein encoded by the BCL2 gene plays a role in regulating apoptosis. Abnormalities in the function and expression level of the Bcl-2 protein are associated with several cancers. Saliva is a body fluid that can be used for biomedical research because it contains essential biomarkers of the body and genetic material derived from free cells in the oral cavity. This study aims to get primer characterization for the RT-PCR method for the BCL2 gene. We used RNA samples isolated from saliva to optimize the primers' annealing temperature, concentration, and combination pairs. Previous studies produced three primer candidates, i.e., primers A, B, and C, used in this research. The optimization results showed that primer C was the best primer to be used in the real-time PCR of this study. The optimal annealing temperature used was 60.3°C with a primer concentration of 400 nM. This study also shows the potential of saliva as a material for biomedical studies on the BCL2 gene. The results of the primer characterization resulting from this research are the first step in establishing the in-house RT-PCR method. The validation research will use a larger sample to validate this method.