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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 ...
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.
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]
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 rpoB gene encodes the β subunit of bacterial RNA polymerase and the homologous plastid-encoded RNA polymerase (PEP). It codes for 1342 amino acids in E. coli, making it the second-largest polypeptide in the bacterial cell. [1] It is targeted by the rifamycin family of antibacterials, such as rifampin. [2]
RNA polymerase (purple) unwinding the DNA double helix. It uses one strand (darker orange) as a template to create the single-stranded messenger RNA (green). In molecular biology , RNA polymerase (abbreviated RNAP or RNApol ), or more specifically DNA-directed/dependent RNA polymerase ( DdRP ), is an enzyme that catalyzes the chemical reactions ...
The Klenow fragment, derived from the original DNA Polymerase I from E. coli, was the first enzyme used in PCR. Because of its lack of stability at high temperature, it needs be replenished during each cycle, and therefore is not commonly used in PCR. The bacteriophage T4 DNA polymerase (family A) was also initially used in PCR. It has a higher ...
A DNA polymerase is a member of a family of enzymes that catalyze the synthesis of DNA molecules from nucleoside triphosphates, the molecular precursors of DNA.These enzymes are essential for DNA replication and usually work in groups to create two identical DNA duplexes from a single original DNA duplex.