Trace nucleic acid releasing agent based on RT-RPA method as well as preparation method and application of trace nucleic acid releasing agent

Abstract

The invention relates to a trace nucleic acid releasing agent based on an RT-RPA method. The trace nucleic acid releasing agent comprises TritonX-100, Tween-20, guanidine isothiocyanate, ethanol, isoamyl alcohol and water. The trace nucleic acid releasing agent provided by the invention has the advantages that the required raw materials are low in price, easy to obtain, and harmless to the body. The trace nucleic acid releasing agent can rapidly destroy a pathogen coat protein structure and release nucleic acid, can greatly shorten the nucleic acid extraction and purification time, does not need complex operation steps, and is simple and convenient to operate. The trace nucleic acid releasing agent disclosed by the invention needs a small amount of samples, and can be used for detecting trace precious samples. The trace nucleic acid releasing agent disclosed by the invention can be used for directly extracting nucleic acid of samples such as serum, whole blood, plasma, throat swab, excrement and the like, complex steps such as heating, centrifuging and the like are not needed, only one sample adding process is needed, and the nucleic acid extraction and release process can be completed within 30 minutes.

Description

technical field [0001] The invention relates to a trace nucleic acid releasing agent based on RT-RPA method, its preparation method and application, and belongs to the technical field of biological reagents. Background technique [0002] Nucleic acid extraction technology is the basis of molecular biology research, and has been widely used in pathogenic microorganism detection, environmental microorganism detection, food safety detection and other fields. Traditional nucleic acid extraction physical methods include boiling method, magnetic bead method, ultrasonic method, etc.; chemical methods include surfactants, alkali lysis, etc.; biological methods mainly include enzymatic methods, etc. At present, the main problems in the extraction of nucleic acid release agents are incomplete cell lysis, resulting in incomplete release of nucleic acid, unstable nucleic acid, especially RNA, which is easily degraded by temperature, enzymes and other factors; Substances for nucleic aci...

AI Technical Summary

Problems solved by technology

[0004] At present, nucleic acid extraction technology is developing rapidly, but there are still some shortcomings. For example, although the phenol-chloroform extraction method uses common laboratory reagents, it is relatively cheap to do

However, reagents such as phenol and chloroform are used, which are highly toxic. Long-term operation has a great impact on the health of personnel, and repeated extractions are required. The recovery rate of nucleic acids is low and the loss is large.

Although the magnetic bead method is safe and non-toxic, it is expensive

Although the spin column method is beneficial to RNA protection, it requires more samples, more consumables, and repeated centrifugation, which is not convenient for high-throughput and automated operation

[0005] The traditional PCR method takes a long time and requires multiple steps such as denaturation, annealing, and extension. Usually, it takes 1-2 hours to complete nucleic acid detection

Images