Journal of Biochemistry and Biotechnology

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Editorial - Journal of Biochemistry and Biotechnology (2021) Volume 4, Issue 1

Polymerase Chain Reaction.

Marilee Gaetano*

Department of Immunology, University of Michigan, United States

Corresponding Author:
Marilee Gaetano
Department of Immunology
University of Michigan
United States
E-mail:
[email protected]

Accepted date: January 20, 2021

Citation: Gaetano M. Polymerase Chain Reaction. J Biochem Biotech. 2021;4(1):8

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Abstract

Polymerase chain reaction (PCR) could also be a way widely used to rapidly make millions to billions of copies of a specific DNA sample, allowing scientists to need a very small sample of DNA and amplify it to an outsized enough amount to review intimately. Polymerase chain reaction, or PCR, may be a technique to form many copies of a selected DNA region in vitro (in a tube instead of an organism). PCR relies on a thermostable DNA polymerase, Taq polymerase, and requires DNA primers designed specifically for the DNA region of interest.

Introduction

Polymerase chain reaction (PCR) could also be a way widely used to rapidly make millions to billions of copies of a specific DNA sample, allowing scientists to need a very small sample of DNA and amplify it to an outsized enough amount to review intimately.

Polymerase chain reaction, or PCR, may be a technique to form many copies of a selected DNA region in vitro (in a tube instead of an organism). PCR relies on a thermostable DNA polymerase, Taq polymerase, and requires DNA primers designed specifically for the DNA region of interest.

Polymerase chain reaction

The polymerase chain reaction has been elaborated in some ways since its introduction and is now commonly used for an honest kind of applications including genotyping, cloning, mutation detection, sequencing, microarrays, forensics, and paternity testing. Typically, a PCR is a three-step reaction [1].

The number of copies doubles after each cycle. Usually 25 to 30 cycles produce a sufficient amount of DNA. In the original PCR procedure, one problem was that the DNA polymerase had to be replenished after every cycle because it isn't stable at the high temperatures needed for denaturation.

The principle of enzymatic replication of the nucleic acids. This method has within the field of biology an irreplaceable role and constitutes one among the essential methods for DNA analysis. It has been widely wont to detect and quantify pathogenic microorganisms that cause various infectious diseases including some arboviruses, STIs, and bacterial infection [2].

Amplification is achieved by a series of three steps: (1) denaturation, in which double-stranded DNA templates are heated to separate the strands; (2) annealing, in which short DNA molecules called primers bind to flanking regions of the target DNA; and (3) extension, during which DNA polymerase extends the 3′ end of every primer along the template strands. These steps are repeated (“cycled”) 25–35 times to exponentially produce exact copies of the target DNA.

PCR consists of a series of 20–40 repeated temperature changes, called thermal cycles, with each cycle commonly consisting of two or three discrete temperature steps (see figure below). The cycling is often preceded by a single temperature step at a very high temperature (>90 °C (194 °F)), and followed by one hold at the end for final product extension or brief storage.

The temperatures used and therefore the length of your time they're applied in each cycle depend upon a spread of parameters, including the enzyme used for DNA synthesis, the concentration of bivalent ions and dNTPs within the reaction, and therefore the melting temperature (Tm) of the primers [3].

PCR makes it possible to get, by in vitro replication, multiple copies of a DNA fragment from an extract. Matrix DNA are often genomic DNA also as complementary DNA obtained by RT-PCR from a messenger RNA extract (poly-A RNA), or maybe mitochondrial DNA. It is a way for obtaining large amounts of a selected DNA sequence from a DNA sample. This amplification is predicated on the replication of a doublestranded DNA template. It is weakened into three phases: a denaturation phase, a hybridization phase with primers, and an elongation phase. The products of each synthesis step serve as a template for the following steps, thus exponential amplification is achieved.

References

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