RAA (recombinase-mediated strand replacement nucleic acid amplification) - A breakthrough thermostatic alternative
In vitro nucleic acid rapid amplification technology is a technology that enables rapid and rapid amplification of trace nucleic acids in vitro. Recombinase-aid amplification (RAA) technology is a constant temperature in vitro rapid amplification nucleic acid technology developed on the basis of existing in vitro nucleic acid amplification principles. The RAA method utilizes recombinase, single-stranded binding protein, and DNA polymerase to replace the thermal cycle melting process of traditional PCR, while achieving rapid amplification of nucleic acids at a constant temperature of 37°C. It is expected to replace traditional thermocycling PCR in the near future.
Principles of RAA Technology
Recombinase-mediated strand replacement nucleic acid amplification technology (RAA technology) is a technique for rapid amplification of constant-temperature nucleic acids using recombinant enzymes obtained from bacteria or fungi. The recombinase can be tightly bound to the primer DNA at room temperature. The polymer forms the enzyme and the primer, when the primer searches the template DNA for a complementary sequence that exactly matches it. With the help of the single-stranded DNA binding protein, this opens the double-stranded structure of the template DNA. The DNA polymerase while under the action, new complementary strands of DNA are formed and the amplification products increase exponentially.
The use of fluorescent probe labels allows for the quantification of the results of timed analysis. Usually, fluorescent detection results can be obtained within 5-15 minutes.
RAA technology advantages:
Simple Operation: The operation is simple and the sample can be tested at the molecular level without the need of a professional.
Rapid Testing：5-15min for test results.
Accurate Results: High sensitivity, specificity, and accurate identification of target gene fragments, so the results obtained are accurate.
Portable Equipment: The equipment is portable, able to cope with various harsh environments, and is law abiding.
Room Temperature Reaction: Constant temperature reaction at 30-42°C, so there is no need to buy any thermal cycler or other expensive equipment.
Freeze-drying Process: Using advanced freeze-drying technology, it can be made into dry powder reagent that can be directly tested in tranportable machines. Operation is more convenient, but also avoids losses caused by repeated freezing and thawing.
Technical Platform Comparison
RAA Technology Application
The kit reaction components have been mixed and lyophilized to a dry powder state. Only the A, B Buffer and the extracted test sample DNA need to be added during use. The operation is simple and easy.
In theory, the RAA technique can be used for multiplex detection, that is, adding two or more pairs of primers to the same RAA reaction system and simultaneously amplifying the RAA reaction of multiple nucleic acid fragments. Multiple RAA technology has the advantages of systematization, low cost, etc. Multiple detection standards can be detected simultaneously in the same reaction tube, which will greatly save the detection reagents.
RAA Technology Application Area
RAA technology has a wide range of applications, including detection of harmful microorganisms in food safety testing, detection of animal-derived components, detection of genetically modified organisms, detection of aquatic diseases, and detection of animal diseases.
Nucleic Acid Test Revolution
RAA technology is a revolution in nucleic acid testing, due to its high sensitivity, specificity, accurate results, rapid detection, low equipment requirements, and ease of operation.