One-step Universal Green Fast RT-qPCR Kit

Quantitative PCR (qPCR), also known as Real-time PCR, is a highly versatile and precise technique for analyzing gene expression. According to the methodology, qPCR can be divided into two categories: dye-based methods and probe-based methods. Among these, dye-based methods are more universal, convenient, and cost-effective. Dye-based qPCR indirectly measures DNA amplification during each PCR cycle by monitoring the fluorescence emitted by dyes that bind specifically to double-stranded DNA (dsDNA). When the fluorescence signal detected at a certain time point significantly exceeds the background, the Ct value (also known as Cq value) can be determined. The obtained Ct values can be used to evaluate the relative abundance of the target gene or to calculate absolute quantities based on an appropriate standard curve.

The One-step Universal Green Fast RT-qPCR Kit is a new type of product specially designed for rapid real-time PCR. It enhances durability and extension speed, and has improvements in signal-to-noise ratio (fluorescence), cyclic domain value (Ct), linearity and sensitivity. This product utilizes the real-time fluorescence signal of the dsDNA-binding dye SYBR Green I to monitor DNA amplification after each PCR cycle, enabling dye-based real-time quantitative analysis of target RNA sequences. The SYBR Green Fast RT-qPCR Enzyme Mix in this kit contains antibody-mediated hot-start Taq DNA polymerase and a novel reverse transcriptase with high thermal stability. The recommended reverse transcription temperature is 55°C, but it can be adjusted between 50°C-55°C. The 2×SYBR Green Fast RT-qPCR Reaction Buffer contains dNTPs, SYBR Green I, and all necessary buffer components. Additionally, the premix includes a unique inert reference dye compatible with various qPCR instruments, including those requiring high or low ROX reference signals. Furthermore, the premix employs a thermolabile UDG+dUTP anti-contamination system, ensuring the accuracy of qPCR amplification results and reducing false-positive results caused by aerosol contamination. This kit also contains a non-fluorescent visible tracking dye, facilitating the visualization of reaction setup without interfering with real-time detection.