One Step PrimeScript III RT-qPCR Mix is a dedicated reagent for one-step real-time, probe-based RT-qPCR (using the 5’ nuclease method). This 2X premix does not freeze at its storage temperature of –20℃, so a reaction can be started simply by adding the template sample, primer, and a probe for detecting the desired target. The quick and simple protocol allows the reverse transcription and qPCR reactions to be performed in the same tube. The reverse transcription reaction uses the novel PrimeScript III RTase, which displays increased heat tolerance (up to 55℃) while maintaining the specificity and extensibility of PrimeScript RTase. This allows cDNA synthesis from RNA with a more complex secondary structure. After cDNA synthesis, TaKaRa Taq HS performs highly specific and efficient PCR amplification, while the fluorescence emitted by the probe is detected in real-time. One Step PrimeScript III RT-qPCR Mix is also highly resistant to a wide variety of inhibitory substances in blood and soil, allowing stable one-step real-time RT-qPCR to be performed on a wide range of samples. This product can be used for various applications such as gene expression, RNA virus detection, etc.