RT-PCR+(Reverse+Transcriptase+Polymerase+Chain+Reaction


 * Basic Description **

Reverse Transcriptase Polymerase Chain Reaction is a technique commonly used in cell biology/genetics, similar to traditional PCR except it uses mRNA instead of DNA to amplify cDNA. This technique is abbreviated RT-PCR, not to be confused with Real-Time PCR, another variant of the Polymerase chain reaction. To carry out an RT-PCR, you need the mRNA segments you wish to amplify, DNA primers, Reverse Transcriptase, Taq DNA Polymerase, dNTPs, and a buffer solution to facilitate the reaction. The only difference (as opposed to PCR) in the reaction is that Reverse Transcriptase makes complementary DNA from mRNA, the rest is the same (Taq DNA Polymerase is still used to amplify over a billion strands of cDNA).

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 * Purpose of Technique **

RT-PCR is used to reverse transcribe mRNA to cDNA and then amplify this result. There are several reasons why researchers would use this opposed to PCR. Since mRNA is the message that is sent for translation, RT-PCR can give us a measurement of gene expression that PCR cannot. The cDNA produced does not contain the introns that DNA does; therefore RT-PCR can provide us with genes that may be inserted into prokaryotes (which cannot and do not splice the introns out). This technique is also used for diagnosing genetic diseases as well as studying certain viruses whose genetic information are exclusively composed of RNA.


 * Origin and History **

Reverse transcriptase was discovered and isolated by Howard Tenim and David Baltimore in 1970. The discovery was very popular at the time because it was thought to be backwards in regards to the central dogma of biology [3]. The 1983 discovery of the PCR technique is attributed to Karl Mullis. Both of these discoveries were awarded with a Nobel Prize. The first person(s) to use RT-PCR is not clear, yet the possibility of this technique seems to go hand-hand with standard PCR and was probably tested shortly after the 1983 discovery.

**Recent Research**

-In a 2004 article, “Transcriptome Profiling of the Response of Arabidopsis Suspension Culture Cells to Suc Starvation” GeneChip analysis is used to measure gene expression in the plant response to nutrient stress. RT-PCR and RNA blots are used to verify the GeneChip expression data. RT-PCR reinforces the results of newer, more cutting-edge technology as to which particular plant genes were up-regulated or down regulated (Contento, Kim, & Bassham, 2004).

- In a 2012 study, “Yield of Sputum for Viral Detection by Reverse Transcriptase PCR in Adults Hospitalized with Respiratory Illness” Novel ways of testing for viral respiratory illnesses are explored. The article states that “viral infection diagnostics have been revolutionized by the development of RT-PCR” and the research performed argues and provides data for the use of RT-PCR assays on sputum samples in detecting these particular viruses (Falsey, Formica, & Walsh, 2012).

- In a 2011 study, “Simultaneous Detection of Rift Valley Fever, Bluetongue, Rinderpest, and Peste des Petits Ruminants Viruses by a Single-Tube Multiplex Reverse Transcriptase-PCR Assay Using a Dual-Priming Oligonucleotide System” the authors discover a novel RT-PCR assay to detect a variety of animal diseases that have a large economic impact. The paper is an example of a way RT-PCR has been tweaked and altered for detection of diseases. The findings of the experiment are simply that the assay works and that they should be implemented (Yeh et. al, 2011).

__ References __ []
 * 1) Central Dogma Reversed". Nature 226 (5252): 1198–1199. 1970. doi:10.1038/2261198a0
 * 2) []