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제품 설명
Hifair™ V2 Multiplex One Step RT-qPCR Probe Kit (UDG Plus, GC enhancer plus)
제품 번호
16630ES
제품 설명
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 HifairTM Reverse Transcriptase and quantitatively amplified by UNICONTM 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.
Components No. | Name | 16630ES60 (100T) | 16630ES80 (1,000T) |
16630-A | 5×HifairTM MP Buffer | 500 μL | 5 mL |
16630-B | HifairTM Enzyme Mix | 100 μL | 1 mL |
16630-C | GC-Enhancer | 500 μL | 5 mL |
[Note]: 1) 5×HifairTM MP Buffer is the abbreviation of HifairTM V2 Multiplex One Step RT-qPCR Probe Buffer, which includes dNTPs, dUTP, Mg2+, stabilizers and more.
2) HifairTM Enzyme Mix mainly contains HifairTM reverse transcriptase, UNICONTM 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
This product should be stored at -25~-15℃ for one year.
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.
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