AccuPower® Dual-HotStart™ RT-qPCR PreMix is a one-step RT-PCR product that applied Pyro-Hotstart RT technology and Hotstart PCR technology to substantially improve the problems of non-specific reverse reaction and enhance the sensitivity to effectively undergo reverse transcription of template RNA even if it is present in a miniscule amount. Moreover, this product is compatible with comprehensive types of devices, including Exicycler™ 96 and those from other companies, by providing optimized tube and plates, along with a 2X master mix type.

Dual-HotStart™ Technology

• 1st hotstart reaction: Hostart RT
  It uses a unique enzyme-mediated HotStart method that provides robust, sensitive, and reliable cDNA synthesis results. Bioneer's RocketScript™ reverse transcriptase is completely inhibited by pyrophosphate at temperatures below 50°C. However, RocketScript™ reverse transceiptase becomes fully active at temperatures above 50°C via pyrophosphate hydrolysis with a thermostable pyrophosphatease. This prevents the formation of mis-primed products and primer-dimers during the reaction set up process resulting in improved specificity of cDNA systhesis.

• 2nd hotstart reaction: HotStart PCR & qPCR
  Bioneer's HotStart Taq Polymerase provides superior priming accuracy and specificity that cannot be achieved with other enzymes. You will use less enzyme per reaction, save money, and get higher sensitivity than with other HotStart enzymes.

Features and Benefits

• Specificity
  The world's first Dual-HotStart™ RT-qPCR reaction using PyroHotStart RT reaction and HotStart PCR optimized to detect only the target gene precisely.
• Sensitivity
  Detection of miniscule amounts of the target templates in a high concentration of RNA samples unable to be done with convientional methods with a wide dynamic range of 10 logs up to 10 ~ 1010 copies.
• Multiplex
  Compatible with many kinds of fluorescent dye (probes) to detect various kinds of target genes.
• Comprehensive template RNA detection
  RocketScript™ RTase included for undergoing RT reaction at high temperature and gaining RT-qPCR results from strong secondary template RNA structures
• Compatibility
  Excellent reactivity towards template RNA allowing accurate RT-qPCR results from wide choices of samples such as from blood and soils continaing various PCR inhibitors
• Convenience
  All reactants necessary for r RocketScript™ RTase, HotStart Taq DNA polymerase and cDNA synthesis and qPCR included in each tubes to readily start the one-step RT-qPCR by adding template DNA, probe & primer for target gene, and D.W.
• Stability
  Stabilizer included in the Real-Time PCR reaction mixture, being more stable the solution type products
• Reproducibility
  Mass production under ISO 9001 quality system allowing minimized deviation between lots and reproducible results even for replicates tested repeatedly under the same conditions and variables

Application

• Gene expression profiling
• Target RNA quantification
• Microbial detection
• Viral/bacterial pathogen load determination

Experimental data

Figure. High sensitivity of AccuPower® Dual-HotStart™ RT-qPCR PreMix. Experiment with HIV target. 10 fold serial dilution of Template RNA (1010 copies ~ 10 copies spiked in Human Total RNA).