2.6. Detection of Amplicon
After completion of real-time PCR, the amplified products were confirmed by gel electrophoresis. Five mL of each amplicon were loaded on 2% agarose gel containing ethidium bromide (EtBr) and SYBR® Gold nucleic acid gel stain (Invitrogen Life Technologies, Eugene, Oregon, USA). The gel electrophoresis was performed at 25 V for 30 min and was visualized by Safe Imager TM (Invitrogen Detection Technologies, Eugene, Oregon, USA).