Probe-based qPCR is favored by experimenters because of its good specificity and multiplex detection and has a wide range of applications in the fields of microbial detection, virus detection, and genotyping. Although probe-based qPCR is much more accurate than dye-based detection, there are still two experimental interferences:
First, if there are PCR product residues or aerosol contamination in the laboratory, the probe-based qPCR may still have the problem of false positives. At this time, it is necessary to introduce a dUTP/UDG anti-contamination system into the qPCR Mix and use UDG enzyme to digest and degrade U-containing DNA molecules to avoid non-specific amplification.
Second, the qPCR amplification effect is also affected by the quality of nucleic acid extracted upstream. The residues of impurities such as proteins and ions will inhibit qPCR amplification. Especially for clinical and veterinary testing, it is difficult to completely remove impurities in samples. At this time, it is necessary to adopt a downstream detection system with strong stress resistance to avoid the phenomenon of low amplification efficiency or ineffective amplification.
In response to these two problems, Genfine has specially launched a master mix for multiplex qPCR that is suitable for DNA templates with strong resistance to stress and contains UDG enzyme: Flash Robust U+qPCR Probe Master Mix. This product is based on the new hot-start polymerase of Flash Robust HotStart DNA Polymerase, combined with double antibody blocking technology and dUTP/UDG anti-pollution system, which can achieve excellent amplification performance and detection sensitivity. At the same time, it has a wide range of compatibility for template type, template GC content and primer Tm value, and has strong resistance to various impurities, which is suitable for various detection scenarios.
▶Excellent amplification performance: high-quality hot-start polymerase is used to ensure high amplification efficiency, strong specificity and good sensitivity.
▶Resistance to Contaminants: high impurity tolerance, especially suitable for direct blood amplification or sample amplification with PCR inhibitors.
▶Support multiple detections: The optimized buffer system can be used for the simultaneous detection of more than 2 target genes.
▶ Equipped with dUTP/UDG anti-pollution system: it can ensure high accuracy and repeatability of quantitative results, and the data is true and reliable.
▶Simple operation: the premix solution simplifies the experimental operation. When configuring the reaction system, you only need to add primers, probes, templates and water.
▶Support rapid procedure: Compatible with rapid amplification, which can greatly improve the detection speed.
1.Excellent detection sensitivity
Using the ASF plasmid as the template, the detection of each gradient sample was carried out. The results showed that A206 has an excellent linear relationship in a wide range of template amounts and can detect single-digit copies of the template to be tested.
2. Resistance to Contaminants
It has good tolerance to both 20% blood and 20% lysate and is better than competing products.
3. Excellent multiple detection capability
Compared with competing products, A206 has excellent amplification efficiency and excellent multiplex detection capability.
4. Stable removal ability
The PCR product of the positive sample (Ct Mean 23.7) was used as the template, and A206 was used for amplification. After digestion and removal by UDG enzyme, the Ct values of the 8 replicate wells were all below 35, indicating that the PCR products in each well occurred. different degrees of degradation. At the same time, A206 was kept at 4°C for 15 days, and it still maintained good scavenging ability.
5. Rapid amplification procedure
The extension rate can reach 1s/kb, and it supports fast programs, which can greatly improve the detection speed.
Note: ROX Reference Dye can be selected according to different models of qPCR instruments.
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