One-step Universal Probe RT-qPCR 4×Mix

Quantitative PCR (qPCR, also known as Real-time PCR) is a highly versatile and precise technique for analyzing gene expression. According to the principle, qPCR can be divided into dye-based and probe-based methods. Commonly used probes include TaqMan probes, molecular beacons, dual-hybridization probes, MGB probes, etc. Among these, TaqMan probes are the most classical and widely used. TaqMan probes specifically hybridize to the target sequence and are labeled with a fluorescent reporter dye at the 5' end and a quencher dye at the 3' end. When the probe is intact, the fluorescence of the reporter dye is quenched due to Fluorescence Resonance Energy Transfer (FRET). During primer-mediated extension, the 5'→3' exonuclease activity of Taq polymerase cleaves the probe bound to the target sequence, separating the fluorescent reporter dye from the quencher dye. Once separated by more than 10 nm, the reporter dye emits fluorescence upon excitation. With each PCR cycle, more fluorescent reporter dyes are released, and the fluorescence intensity is proportional to the amount of newly synthesized target fragments, enabling detection. Common fluorescent reporter dyes at the 5' end include FAM, HEX, TET, JOE, NED, Cy3, TAMRA, Cy5, Cal Fluor Gold540, Cal Fluor Orange560, etc. Common quencher dyes at the 3' end include BHQ1, BHQ2, BHQ3, TAMRA, Dabcyl, NFQ-MGB, etc. When designing probes, select appropriate dyes according to the qPCR instrument model, target gene, and experimental type.

One-step Universal Probe RT-qPCR 4×Mix is a one-step real-time fluorescent quantitative PCR premix based on the probe method, designed for detecting target RNA. Its working principle is as follows: first, reverse transcriptase converts RNA into cDNA; subsequently, DNA polymerase amplifies the cDNA. The entire process is completed in a single tube. Additionally, this product employs a Thermolabile UDG+dUTP anti-contamination system, ensuring accurate qPCR amplification results and reducing false positives caused by aerosol contamination. This product contains specially formulated ROX reference dye, suitable for all qPCR instrument platforms without adjusting ROX concentration, ensuring consistent and reliable results. It also contains a non-fluorescent visible tracking dye for easy observation during reaction setup, which does not interfere with real-time detection.

One-step Universal Probe RT-qPCR 4×Mix is a convenient 4× premix, requiring only the addition of primers, probes, and templates. It is easy to operate, compatible with fast protocols, significantly reducing detection time. It provides high accuracy and reproducibility for target gene quantification across a wide dynamic range. This product is suitable for pathogen detection, RNA virus detection, gene expression analysis, and other applications.