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RT-PCR. 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.
A reverse transcriptase (RT) is an enzyme used to convert RNA genome to DNA, a process termed reverse transcription.Reverse transcriptases are used by viruses such as HIV and hepatitis B to replicate their genomes, by retrotransposon mobile genetic elements to proliferate within the host genome, and by eukaryotic cells to extend the telomeres at the ends of their linear chromosomes.
Template-switching polymerase chain reaction (TS-PCR) is a method of reverse transcription and polymerase chain reaction (PCR) amplification that relies on a natural PCR primer sequence at the polyadenylation site, also known as the poly(A) tail, and adds a second primer through the activity of murine leukemia virus reverse transcriptase. [1]
The protocols for 5' or 3' RACES differ slightly. 5' RACE-PCR begins using mRNA as a template for a first round of cDNA synthesis (or reverse transcription) reaction using an anti-sense (reverse) oligonucleotide primer that recognizes a known sequence in the middle of the gene of interest; the primer is called a gene specific primer (GSP). The ...
The outer primers(F3 and B3) anneal to the template strand and help the reaction to proceed. As in the case of RT-PCR, the RT-LAMP procedure starts by making DNA from the sample RNA. This conversion is made by a reverse transcriptase, an enzyme derived from retroviruses capable of making such a conversion. [15]
Vectorette PCR has been found to be beneficial when it comes to the characterization of these intronic sequences when they are found to be next to known sequences. [21] cDNA or complementary DNA is a DNA sequence which is complementary to the RNA that is the template when synthesizing DNA during the reverse transcriptase process. [22]
RT-PCR (or Reverse Transcription PCR) is used to reverse-transcribe and amplify RNA to cDNA. PCR is preceded by a reaction using reverse transcriptase , an enzyme that converts RNA into cDNA. The two reactions may be combined in a tube, with the initial heating step of PCR being used to inactivate the transcriptase. [ 4 ]
Transcription-mediated amplification (TMA) is an isothermal (performed at constant temperature), single-tube nucleic acid amplification system utilizing two enzymes, RNA polymerase and reverse transcriptase. "Amplification" means creating many more copies of a strand of nucleic acid than was present at first, in order to readily detect it or ...
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