Nucleic acid amplification system and freeze-drying protective agent thereof

Abstract

The invention discloses a nucleic acid amplification system and a freeze-drying protective agent thereof. The nucleic acid amplification system comprises a buffer solution, an enzyme mixing solution, a nucleic acid amplification reaction solution and the low-temperature freeze-drying protective agent. The buffer solution further comprises one or more of dNTP, Mgcl2, Thris-Hcl and KCl. The enzyme mixing solution further comprises one or more of RNA reverse transcriptase, RAN inhibitor and DNA polymerase. A preparation method for the freeze-drying protective agent applied to the nucleic acid amplification system comprises the following step that the components are mixed to be uniform according to the adopted concentration ratio to obtain the freeze-drying protective agent. An RNA / DNA fluorescent quantitation nucleic acid diagnostic reagent system can be detected in a freeze-drying mode, freeze drying can be achieved, the detection purpose can be achieved only through one-step hydration, and the purposes of normal temperature transportation and storage which cannot be achieved by a nucleic acid diagnostic reagent at the present stage can be achieved.

Description

technical field [0001] The invention relates to the technical field of molecular biology, in particular to a freeze-drying protective agent applied to a nucleic acid amplification system. Background technique [0002] Polymerase chain reaction (PCR) technology has been used in medical testing and clinical testing since 1989 based on its advantages of rapidity, simplicity, sensitivity, and specificity, and soon became one of the main detection methods in medicine and diagnostics. [0003] DNA nucleic acid amplification is a molecular biology technique used to amplify specific DNA fragments. Using DNA denaturation at high temperature in vitro will become a single strand. At low temperature, primers and single strands will combine according to the principle of base complementary pairing, and then adjust the temperature to the optimum reaction temperature of DNA polymerase. DNA polymerase will follow the phosphoric acid to the five-carbon sugar Complementary strands are synthes...

AI Technical Summary

Problems solved by technology

[0006] Nucleic acid diagnosis is widely used in the detection of various pathogens, but nucleic acid amplification reaction reagents are unstable when stored at room temperature, and become invalid in a few weeks; when stored at 2-8°C, the storage period is only about three months; Although the storage period at -20°C can be as long as one year, it must be repeatedly frozen and thawed each time it is used, which also has a great impact on the activity of the reagent, and the components cannot be mixed together. Trouble and other disadvantages, the long-term storage and long-distance transportation of the kit will be greatly restricted, and the sensitivity of the diagnostic reagent will be reduced or even completely invalid due to improper storage temperature, which will eventually lead to the untimely detection of the disease and cause the epidemic

[0007] At present, nucleic acid amplification reaction reagents on the market lack methods that can be stored at 4°C or room temperature for a long time, which also limits the market for nucleic acid diagnostic reagent products

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