Overview of the PCR and its Applications 1 Overview of the PCR and its Applications By [Name] Course Professor’s Name Institution Location of Institution Date
Overview of the PCR and its Applications 2 Introduction Created in 1983 by Kary Mullis, the polymerase chain reaction (PCR) is presently a typical and regularly key strategy utilized in medicinal and natural research labs for an assortment of utilization. PCR is a quick, economical and basic technique for replicating specific DNA molecules from very small quantities of the DNA material, even in cases where that source DNA is of poor quality. The technology does not use radioisotopes or dangerous synthetic substances making it safe and appropriate for numerous applications. One of the reasons for amplifying DNA is to make various duplicates of a piece of DNA which is extremely rare. For instance, a forensic researcher may need to amplify little pieces of DNA from a crime scene to facilitate investigations. Also, one may need to make comparisons between two or more DNA samples to ascertain which one is more abundant The PCR The chain reactions capitalize on the ability of an enzyme called polymerase to make new DNA strands complementary to the gene of interest. A polymerase will orchestrate a corresponding arrangement of bases to any single strand of DNA as long as it has a double- stranded beginning point. The DNA of interest forms the template strand in which the DNA polymerase adds nucleotides onto it at the 3’-OH group. Therefore, the DNA polymerase requires short single-stranded pieces of DNA called oligonucleotides complementary to the desired DNA or a section of it. These oligonucleotides are referred to as primers which with the activity of the enzyme amplify the specific regions of a DNA strand. In most of these reactions 0.1 to 10-kilo base pairs (kbp) of small pieces of DNA are amplified. However, some other techniques can achieve amplification of up to 40kbp (Tellinghusen and Spiess, 2014, pp.76-82).
Overview of the PCR and its Applications 3 The available substrates influence the amount of the products because as the reaction progresses they get used up leading to product reduction (Carr and Moore, 2012, p.37640). Components of a standard PCR For a standard set-up, the PCR requires various components and chemicals. A template DNA containing a region for amplification together with a DNA polymerase enzyme that will amplify it is required. The enzyme ought to be heat resistant since the reaction uses extreme temperatures. Taq and Pfu polymerase are the most common catalysts used to drive these reactions (Hommelsheim et al., 2014, p.5052). The DNA needs two DNA primers complementary to the double DNA strands which form the anti-sense and sense strands. The primers have to be two for the activity of the DNA polymerase is only initiated when it binds to double strands. The DNA polymerase obtains nucleotides from deoxy-nucleoside triphosphates
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