Dry-powder isothermal amplification detection reagent capable of being transported at normal temperature, and preparation method of detection reagent

Abstract

The invention discloses a dry-powder isothermal amplification detection reagent capable of being transported at normal temperature, and a preparation method of the detection reagent. The detection reagent can be prepared by the following steps: adding a freeze-drying protective additive into a liquid reagent, and adopting a vacuum freeze drying method to freeze-dry components of the liquid isothermal amplification detection reagent to be solid dry powder. The solid reagent can be transported at normal temperature; during reaction, all that is needed is to add a liquid reaction substrate to be dissolved; the reactivity is excellent, the operation is convenient, and the dry-powder isothermal amplification detection reagent can be widely used in on-site quick detection of various pathogenes, food microorganisms, transgenic products and the like.

Description

technical field [0001] The invention relates to a dry-powdered constant-temperature amplification detection reagent that can be transported at normal temperature and a preparation method thereof. Background technique [0002] Loop-mediated isothermal amplification technology is a novel constant-temperature nucleic acid amplification method developed by Japan's Eiken Chemical Co., Ltd. in 2000. It is characterized by designing 4 specific primers for 6 regions of the target gene and utilizing a strand displacement The nucleic acid amplification reaction can be completed by incubating the DNA polymerase for 30-60 minutes under isothermal conditions (about 63°C). Compared with conventional PCR, thermal denaturation of the template, temperature cycling, electrophoresis and ultraviolet observation are not required. The loop-mediated isothermal amplification method is a brand-new nucleic acid amplification method, which has the characteristics of simplicity, rapidity and strong sp...

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Problems solved by technology

[0010] However, these substances require refrigerated storage, especially Bst DNA polymerase needs to be stored at -20°C, and it cannot be frozen and thawed repeatedly. Therefore, it brings great trouble to the transportation of reagents and customer use, and increases the transportation cost. At the same time, a corresponding reaction system must be prepared for each experiment , it is also very inconvenient to use, it is easy to cause errors caused by the operator's operation, and it is easy to cause contamination and cross-contamination between experiments

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