Nucleic acid releasing agent, nucleic acid PCR (polymerase chain reaction) amplification method and PCR amplification kit

Abstract

The invention relates to a nucleic acid releasing agent, a nucleic acid PCR (polymerase chain reaction) amplification method and a PCR amplification kit. The nucleic acid releasing agent comprises Tris-HCl, sodium chloride, potassium chloride, Tween 20, Triton X-100, Nonidet P-40 and strong base; the molar concentration of the Tris-HCl is 0.5-500mM, the molar concentration of the sodium chloride is 20-500mM, the volume percentage of the Tween 20 is 0.1-2%, the volume percentage of the Triton X-100 is 0.1-3%, the volume percentage of the Nonidet P-40 is 0.1-3%, the mass concentration of the potassium chloride is 5-8mg / mL, and the mass concentration of the strong base is 2-50mg / mL. The nucleic acid releasing agent has the advantages that RNA-containing samples can directly release RNAs at the room temperature, and can be directly mixed with PCR reaction liquid to achieve PCR amplification, so that amplification can be completed without complex nucleic acid extraction and purification processes.

Description

technical field [0001] The invention relates to the field of molecular biology, in particular to a nucleic acid releasing agent, a nucleic acid PCR amplification method and a PCR amplification kit. Background technique [0002] PCR (Polymerase Chain Reaction, Polymerase Chain Reaction) is a molecular biology technique used to amplify specific nucleic acid fragments. The purpose of detecting trace amounts of nucleic acid. Among them, the commonly used PCR method in medical diagnosis is mainly the real-time fluorescent quantitative PCR (qPCR) method based on dual fluorescent probes. The targets for in vitro diagnosis using qPCR method mainly include human genomic DNA, DNA viruses, bacteria, fungi and RNA viruses, etc. . However, since the structure of RNA is a single-stranded structure, which is unstable and easy to degrade, the requirements in the sample processing process are very high, and more complicated methods are required for pretreatment and nucleic acid extraction ...

AI Technical Summary

Problems solved by technology

However, since the structure of RNA is a single-stranded structure, which is unstable and easy to degrade, the requirements in the sample processing process are very high, and more complicated methods are required for pretreatment and nucleic acid extraction and purification of the RNA sample to be amplified to obtain pure After the nucleic acid is removed, the test can be carried out to obtain stable results

Images