Polymerase Chain Reaction (PCR) - It Is Now One Of The Most Widely Used Molecular Biology Techniques20/3/2023 The amplification of nucleotide sequences is made possible by the Polymerase Chain Reaction (PCR), a method that has revolutionised molecular biology and other fields. A brief history of PCR and its guiding principles, as well as an examination of its various forms and the particular uses to which they are put, are covered in this article. One of the most often used tests for acquiring a specific DNA or RNA segment is the Polymerase Chain Reaction (PCR). It moves quickly and is very perceptive. It offers a way to isolate that specific DNA segment or gene by amplifying the chosen DNA fragment. The nucleotide sequence at the endpoints of the region you want to amplify must be known to use this strategy. Once that is known, one can start with extremely small amounts of material, like the DNA found in a single human hair, and produce huge volumes of that region. The variety of genes to which it can be applied is huge because to the availability of almost entire or complete sequences of genomes from numerous species. There are several uses for PCR, including diagnostics, forensics, gene isolation, and investigations of gene expression. The strength of Polymerase Chain Reaction (PCR) comes from the exponential growth in DNA produced by repeated cycles of DNA synthesis using primers that flank a specific region, one primer directed at directing synthesis complementary to the top strand and the other primer directed at directing synthesis complementary to the bottom strand. The amount of DNA that is produced in each cycle increases by roughly a factor of two when this is done repeatedly. As a result, with enough cycles, the amount of DNA from the amplified area can be increased by a million-fold. The polymerase chain reaction gets its name from the fact that this exponential increase in abundance resembles a chemical chain reaction. In the first cycle, the reaction template is the only one available, and it is still a significant template in the second cycle. A product with ends determined by primers is created at the conclusion of the second cycle. This is the desired outcome, and it acts as the primary model for the subsequent cycles. Although the original template is still there and can still be utilised, it does not experience the exponential growth seen for the desired product. The discovery of DNA polymerases that were stable at high temperatures significantly improved the simplicity of performing Polymerase Chain Reaction (PCR). They have been identified from bacteria like Thermus aquaticus that thrive in hot springs like those in Yellowstone National Park. Even at the high temperatures required to melt the templates, this bacterium's Taq polymerase will continue to function, and it is active between the melting and annealing temperatures. Several thermostable polymerases have been found that are more accurate than this one, which is fairly error-prone.
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