One Step PrimeScript III RT-qPCR Mix, with UNG 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. Because the premix contains Uracil N-Glycosylase (UNG), which degrades amplification products from the previous reaction, it can prevent carryover contamination even when a specific target is repeatedly tested for the purpose of inspection, etc. 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, with UNG is also highly resistant to a wide variety of inhibitory substances such as heparin (blood) and humic acid (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.