Freeze-drying protective agent, PCR amplification reagent, freeze-drying method and application thereof

Abstract

The invention is applicable to the field of biotechnology diagnosis and detection, and provides a freeze-drying protective agent, a PCR amplification reagent, a freeze-drying method and application thereof. The PCR amplification reagent comprises an amplification reaction system and the freeze-drying protective agent, wherein the freeze-drying protective agent comprises the following components: glycerol, trehalose, sucrose, Tween-80, dextran-40, glycine, histidine, sodium glutamate, a bacteriostatic agent and tert-butyl alcohol. The freeze-drying protective agent provided by the invention canbe prepared together with components required for fluorescent PCR, and can also be matched with any primers and probes required for PCR detection, and freeze-drying is carried out to prepare the PCRamplification reagent; and the PCR amplification reagent can be transported and stored at normal temperature, and only ddH2O is needed to be dissolved and then sample nucleic acid is added when the PCR amplification reagent is used, so that the PCR amplification reagent has the advantage of convenient operation.

Description

technical field [0001] The invention belongs to the field of biotechnology diagnosis and detection, and in particular relates to a freeze-drying protective agent, a PCR amplification reagent and a freeze-drying method and application thereof. Background technique [0002] Real-time fluorescent quantitative PCR (Quantitative Real-time PCR) is a method for detecting the total amount of products after each polymerase chain reaction (PCR) cycle with fluorescent chemicals in DNA amplification reactions. Fluorescent quantitative PCR technology is currently the most commonly used detection method in the field of diagnostic testing, because this method is highly sensitive, specific, simple and fast, and the detection result can be obtained within a few hours. The necessary conditions for the reaction include: a fluorescent PCR reaction system, primers and probes. [0003] Among them, the components of the fluorescent PCR reaction buffer system are usually chemical reagents and wate...

AI Technical Summary

Problems solved by technology

The operation method of this form of product is cumbersome, and it needs to be artificially prepared before use, and the PCR product needs to be frozen at -20°C. First, the kit must be warmed up, and then the reaction buffer and enzyme are mixed evenly according to the recommended ratio of the kit. Put it into a PCR reaction tube, and then add the sample, the more times of freezing and thawing, the easier the enzyme activity is to degrade

Because the amount of enzyme is usually less (≤1μL), in order to stabilize the enzyme, there is glycerol in the enzyme solution, which is relatively viscous, and the preparation error is large, the reaction repeatability is not good, and cross-contamination is also prone to occur.

Moreover, reagents need to be stored at low temperature (below -20°C), and the cost of product storage and transportation is relatively high

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