One Step PrimeScript III RT-qPCR Mix is a 2X master mix for one-step real-time, probe-based RT-qPCR (using the 5′ nuclease method). It contains a novel heat- and inhibitor-resistant reverse transcriptase, PrimeScript III RTase, in combination with an efficient, specific hot-start PCR enzyme, TaKaRa Taq HS, and an optimized reaction buffer. A quick and simple protocol allows the reverse transcription and qPCR reactions to be performed in the same tube, reducing the risk of contamination.

PrimeScript III RTase displays increased heat tolerance (up to 55°C) while maintaining the specificity and robustness of PrimeScript RT. This allows cDNA synthesis from RNA with a more complex secondary structure. It 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 variety of samples. While One Step PrimeScript III RT-qPCR Mix often works well with crude RNA extracts, using a highly pure RNA sample is recommended if the target sequence is not detectable or if more accurate, stable analysis results will be needed. It can be used for various applications such as gene expression and RNA virus detection, and it is well-suited for multiplex real-time PCR.