Dye based Real-time Quantitative PCR (qPCR)
 
SupRealQ Ultra Hunter SYBR qPCR Master Mix (U+), SKU: Q713
Violet Trace, Easy Tracking: Stable Amplification, Boundless Potential. 
 
Features
Low Expression, High Precision: Designed for low-abundance targets, it captures low-template, low-concentration, and low-input signals with superior stability, outperforming similar products.
 
Effortless Handling of Complex Templates: Handles degraded samples and high-GC templates with ease, delivering reliable results even in challenging experimental scenarios.
 
Smart Anti-Contamination, Worry-Free Experiments: Equipped with a dUTP/Heat-labile UDG system, it effectively eliminates contaminants at room temperature, ensuring cleaner and more reliable experiments.
 
Pre-Mixed Technology, Unmatched Stability: Features a fully premixed formula, maintaining consistent performance even after mixing with primers and templates and storing for 72 hours, offering unparalleled flexibility and stability.
 
Components
See Table above.
 
Performance
Compatibility with Complex Samples
Using the BioSmart platform, we screened core enzymes for high amplification performance and combined them with patented dual-species antibody blocking technology. This combination ensures stable amplification of complex templates, reduces the need for repeated system adjustments, and improves researchers' experimental efficiency.
 
a. Reliable Detection of Relatively Low-Abundance Genes
Using SupRealQ Ultra Hunter SYBR qPCR Master Mix (U+) (Vazyme #Q713), qPCR reactions were performed on 293T cDNA from different genes under identical conditions. The CT values ​​of some genes were delayed compared to others (as shown in Figure 1 upper panel, for genes 1, 2, and 3), indicating that they are classified as relatively low-abundance genes.
 
Gene 3 was amplified using Vazyme #Q713 and a competitor's dye-based qPCR reagent (Supplier A). The results demonstrated that Vazyme #Q713 demonstrated superior sensitivity for low-abundance genes compared to Supplier A while maintaining high specificity of the amplified product (Figure 1).
 
b. Amplification of Moderately Degraded Samples
Gel electrophoresis of RNA extracted from pancreatic tissue of Litopenaeus vannamei revealed no distinct bands (Figure 2), indicating degradation. The degraded RNA was reverse transcribed into cDNA and then serially diluted in two-fold increments. Shrimp genes were amplified using Vazyme #Q713 and dye-based qPCR reagents from Supplier A on the ABI QuantStudio 3 System.
 
The results demonstrated that Vazyme #Q713 consistently amplified degraded samples, with amplification efficiencies meeting the 90-110% standard specified by the MIQE (Minimum Information for Publication of Quantitative Real-Time PCR) guidelines (Figure 2).
 
c. Amplification of GC-High and GC-Low Genes
Genes 4 and 5, representing high and low GC content, respectively, were amplified using 293T gDNA as template using Vazyme #Q713 and dye-based qPCR reagents from Supplier A on the ABI QuantStudio 3 System (Figure 3).
 
Results showed that Vazyme #Q713 demonstrated superior sensitivity and specificity compared to Supplier A for both high- and low-GC-content genes.
 
Wide Linear Range
Mouse liver cDNA was serially diluted 10-fold, and then qPCR amplified using Vazyme #Q713 and dye-based qPCR reagents from Supplier A on the ABI QuantStudio 3 System (Figure 4).
 
The results showed that Vazyme #Q713 exhibited 90-110% amplification efficiency over a wide linear range, meeting the robustness standards of the MIQE guidelines and outperforming Supplier A (Figure 4).
 
Efficient Anti-Contamination
Vazyme #Q713 utilizes a dUTP/thermolabile UDG anti-contamination system, effectively removing contaminants from the reaction mixture at room temperature. When the reaction temperature is increased to 50-55°C, the thermolabile UDG is rapidly inactivated, preserving the integrity of the cDNA and ensuring detection sensitivity.
 
To test contaminant removal efficiency, 60 pg and 600 pg of U-containing template were added to the reaction mixture, respectively (Figure 5). The results showed that Vazyme #Q713 achieved contaminant removal efficiencies exceeding 99.99%, effectively ensuring the accuracy of the experimental results.
 
Product Manual
Manual_Q713 V24.1