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In contrast to similar techniques such as polymerase chain reaction and ligase chain reaction, this method involves RNA transcription (via RNA polymerase) and DNA synthesis (via reverse transcriptase) to produce an RNA amplicon (the source or product of amplification) from a target nucleic acid. This technique can be used to target both RNA and ...
Recombinase polymerase amplification (RPA) is a single tube, isothermal alternative to the polymerase chain reaction (PCR). [1] By adding a reverse transcriptase enzyme to an RPA reaction, it can detect RNA as well as DNA , without the need for a separate step to produce cDNA .
Reverse transcription polymerase chain reaction (RT-PCR) is a laboratory technique combining reverse transcription of RNA into DNA (in this context called complementary DNA or cDNA) and amplification of specific DNA targets using polymerase chain reaction (PCR). [1] It is primarily used to measure the amount of a specific RNA.
The method consists of three separate steps: in the first step, the poly-adenylated RNA is hybridised to a DNA oligonucleotide featuring a poly-deoxythymidine sequence at its 5’ end. Klenow polymerase then catalyses elongation of the mRNA’s 3’ end, using the DNA oligonucleotide as a template. This reaction takes place at 25 °C.
RNase H then degrades the RNA template and the other primer binds to the cDNA to form double stranded DNA, which RNA polymerase uses to synthesize copies of RNA. [11] One key aspect of NASBA is that the starting material and end product is always single stranded RNA. That being said, it can be used to amplify DNA, but the DNA must be translated ...
Reverse transcription loop-mediated isothermal amplification (RT-LAMP) is a one step nucleic acid amplification method to multiply specific sequences of RNA. It is used to diagnose infectious disease caused by RNA viruses. [1] It combines LAMP [2] DNA-detection with reverse transcription, making cDNA from RNA before running the reaction. [3]