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, TaKaRaTaq 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.
- 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.
- Contains sufficient reagent for performing 200 reactions in a volume of 25 µl per reaction.
Overview
- Fast, easy one-step RT-qPCR in a single tube using probe-based detection
- Efficient, specific cDNA synthesis and amplification, even from complex RNA templates
- Suitable for a wide variety of samples from different sources, including crude RNA extracts
Applications
- Gene expression
- RNA virus detection
- Multiplex real-time PCR
Instrument compatibility
- Thermal Cycler Dice Real Time System (TaKaRa Cat. #TP800)2
- ABI PRISM 7000/7700/7900HT, 7300/7500 Real-Time PCR System (Applied Biosystems)
- LightCycler (Roche Diagnostics)
- SmartCycler System/SmartCycler II System (Cepheid)
Storage
- –20°C
