One-Step RT-PCR Quantification Assay

A one-step RT-PCR was performed using TaqMan with BHQ quencher probe at 125 nM and 250 nM of primers in a final volume of 20 μL. Five microliters of the extracted RNA was combined with 15 μl of the master mix and the reverse transcription step was performed 95°C for 15 minutes, 60 cycles of 15 seconds at 95°C and 45 seconds at 60°C. All the procedure was performed in Light Cycler^® 2.0 Instrument and data was analyzed with the LightCycler^® Software 4.1 (Roche Diagnostic, Deutschland-Mannheim, Germany). The primers and the probe used are shown in Table 1 [19 (link)–21 (link)].