Fluorescent quantitative PCR reaction fluid and method

Abstract

The invention discloses a fluorescent quantitative PCR reaction fluid and a method; the fluorescent quantitative PCR reaction fluid at least includes DNA polymerase with ordinary excision enzyme activity, a basic fluorescent stabilizer, DNA polymerase enhancer and protection agent. The fluorescent quantitative PCR reaction fluid of the invention is able to obviously improve the 5'-3- excision enzyme activity of the DNA polymerase, so that the ordinary DNA polymerase can hydrolyze Taqman-MGB probe, improve the amplification efficiency and keep strong stability. The fluorescent quantitative PCR reaction fluid has stronger sensitivity in the application of Taqman-MGB probe, the effect is obviously better than that of the prior art; therefore, the fluorescent quantitative PCR reaction fluid has very wide market prospect.

Description

technical field [0001] The invention relates to a fluorescent quantitative PCR reaction solution and method, in particular to a fluorescent quantitative PCR reaction solution and method applied to Taqman-MGB probes, and belongs to the field of molecular biology technology and experimental methods. Background technique [0002] During the amplification of Taqman fluorescent quantitative PCR technology, a specific Taqman fluorescent probe, also known as a hydrolysis probe, is added at the same time as a pair of primers. inactivate the fluorophore. Commonly used fluorescence quenching groups include TAMRA, BHQ1 and MGB. MGB (Minor Groove Binder) is a minor groove binder, which itself is a chemical group that can further form hydrogen bonds with the minor groove in the probe-template DNA duplex, thereby improving the binding stability of the Taqman-MGB probe to the template sex. [0003] Taqman-MGB probes have many special and excellent properties: 1. Taqman-MGB probes are sh...

AI Technical Summary

Problems solved by technology

[0005] However, there are two defects in the Taqman-MGB probe of the prior art: one is that in the process of enzymatic degradation of the MGB probe, a DNA polymerase with high activity of 5'-3' exonuclease is required, and this high exonuclease The active DNA polymerase is monopolized by a few manufacturers, and the cost is very high; the second is that the fluorescent quantitative PCR reaction solution of the Taqman-MGB probe requires a special reaction solution

[0007] Since Taqman-MGB probes have many influencing factors in the process of identifying templates, the fluorescent quantitative PCR reaction solution of ordinary Taqman probes is not suitable for Taqman-MGB probes

In the prior art, there are only 1-2 manufacturers producing special reaction solutions for hydrolyzed Taqman-MGB probes. These products are expensive and not perfect in terms of stability, amplification efficiency and detection sensitivity.

[0008] Therefore, the DNA polymerase and reaction solution suitable for MGB limit the application and promotion of Taqman-MGB probes

Images