The document describes the procedure for preparing a RT-PCR master mix and performing one-step RT-PCR. Key steps include: 1) Preparing a master mix containing 2x RT-PCR Premix, forward and reverse primers, RNA sample, and H2O. 2) Programming the thermal cycler for reverse transcription, PCR activation, denaturation, annealing and data acquisition steps. 3) Placing the PCR tubes in the thermal cycler to start the RT-PCR program.