Hifair™ V2 Multiplex One Step RT-qPCR Probe Kit (UDG Plus)

This kit is a multiplex quantitative PCR kit based on RNA as template. In the process of the experiment, reverse transcription and quantitative PCR were carried out in the same tube, which simplified the experimental operation and reduced the risk of contamination.

In this kit, the first strand cDNA was efficiently synthesized by heat-resistant Hifair^TM Reverse Transcriptase  and quantitatively amplified by UNICON^TM  HotStart Taq DNA Polymerase. The kit mainly contains optimized MP buffer, enzymes mix, GC-Enhancer etc. The buffer solution already contains Mg²⁺ and dNTP. In addition, the factors that can effectively inhibit the  non-specific PCR amplification and improve the amplification efficiency of multiple qPCR reactions are added, which can ensure the amplification efficiency and carry out up to multiple  amplification reaction. The dUTP/UDG system was added to effectively prevent the risk of aerosol contamination.

Features

• Strong versatility - suitable for one-step RT-qPCR detection of nucleic acids from multiple species such as pathogenic microorganisms, human genomes, and plants;
• High sensitivity: The carefully optimized buffer system improves the detection sensitivity of low-concentration templates up to 250 copies/mL;
• dUTP/UDG anti-pollution system: dUTP/ UDG anti-pollution system is introduced to degrade aerosol pollution of products efficiently, reduce false positive, and ensure authentic results;
• Support fast program: compatible with fast program, 30min can produce results.
• Super stability: Stable performance after 14 days at 37°C, long-distance transport, and 10 freeze-thaw cycles.

Components

[Note]: 1) 5×Hifair^TM MP Buffer is the abbreviation of Hifair^TM V2 Multiplex One Step RT-qPCR Probe Buffer, which includes dNTPs, dUTP, Mg2+, stabilizers and more.

2) Hifair^TM Enzyme Mix mainly contains Hifair^TM reverse transcriptase, UNICON^TM HotStart Taq DNA polymeras, RNase inhibitor, and UDGase and so on.

3) GC-Enhancer can decide whether to add to the system according to the situation

Shipping and Storage

This product should be stored at -25~-15℃ for one year.

Application

RT-qPCR Detection; Diagnostic pathogen detection (respiratory tract, intestinal tract, etc.); animal pathogen detection (non-swine fever, etc.); food testing (microorganisms, etc.)

Figures

1. Excellent amplification performance

Figure 1. Amplification efficiency test

16630 linear range results: amplification efficiency meets 90%-110%, and the template range when the slope of the standard curve is 3.3 is 10⁶ copies/μL ~ 10^-2 copies/μL.

There are 8 dilution gradients in total, and the lowest concentration of Supplier A* was not detected.

2. High detection rate

Figure 2. Comparison of amplification performance of RT-qPCR kits from different brands

Amplification of gradient samples using 16630 and similar products from other suppliers in respiratory targets showed that the results showed that 16630ES performed better in terms of detection rate, fluorescence peak increase, Ct value, etc.

3. High sensitivity

Figure 3. Sensitivity comparison test

Using the parainfluenza quadruple primer probe, 20 replicate wells of parainfluenza T7-RNA plasmid were tested, and the results showed that the sensitivity of pathogen quadruple target amplification could reach 0.25 copies/μL.

4. High Inhibitor Tolerance

Figure 4. Inhibitor tolerance testing

16630 and supplier-A fully tolerated 4% ethanol, and 4% whole blood inhibited both to a similar extent.

5. Stability Test

 Figure 5. Thermal accelerated stability testing(A) and Freeze-thaw stability(B) 

(A) Accelerated Stability Test:
Enzyme and buffer components of the single-component 16630 reagent (two-tube format: buffer and enzyme) were stored at 4°C and 37°C for 14 days. Under quadruplex pathogen and human target detection, amplification performance showed no significant difference compared to −20°C storage, indicating qualified stability.

(B) Freeze–Thaw Stability Evaluation:
Amplification performance was evaluated after 10 freeze–thaw cycles using dry ice and −20°C storage conditions. Both pathogen and human quadruplex targets were tested. Results showed qualified stability under both freeze–thaw conditions for all target sets. 

Documents:

Safety Data Sheet

16630_MSDS_HB20250530_EN.PDF

Manuals

16630_Manual_Ver.EN20241114.pdf