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DNA and RNA Isolation Techniques for Non-ExpertsPolymerase Chain Reaction (PCR)

DNA and RNA Isolation Techniques for Non-Experts: Polymerase Chain Reaction (PCR) [Polymerase chain reaction or PCR is a test to detect and amplify DNA (or cDNA) in an organism. In principle, this technique is similar to in vivo transcription and cloning. The isolated DNA and RNA (after converting it into cDNA through reverse transcription) serve as the starting material for any PCR procedure. The three basic steps involved in this process are denaturation of the template (generally ds DNA), annealing of the primers (for specific DNA sequence amplification), and, finally, the extension of the DNA stretch. A combination of different enzymes, buffers, and temperatures in program-controlled equipment (known as PCR machine) helps amplify DNA. The final product (known as amplicon) can be detected at the end through agarose gel electrophoresis or analyzed through different detection methods on a real-time basis. PCR has a broad range of applications, including basic research, medical diagnostics, forensics, and agriculture.] http://www.deepdyve.com/assets/images/DeepDyve-Logo-lg.png

DNA and RNA Isolation Techniques for Non-ExpertsPolymerase Chain Reaction (PCR)

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Publisher
Springer International Publishing
Copyright
© The Editor(s) (if applicable) and The Author(s), under exclusive license to Springer Nature Switzerland AG 2022
ISBN
978-3-030-94229-8
Pages
157 –163
DOI
10.1007/978-3-030-94230-4_20
Publisher site
See Chapter on Publisher Site

Abstract

[Polymerase chain reaction or PCR is a test to detect and amplify DNA (or cDNA) in an organism. In principle, this technique is similar to in vivo transcription and cloning. The isolated DNA and RNA (after converting it into cDNA through reverse transcription) serve as the starting material for any PCR procedure. The three basic steps involved in this process are denaturation of the template (generally ds DNA), annealing of the primers (for specific DNA sequence amplification), and, finally, the extension of the DNA stretch. A combination of different enzymes, buffers, and temperatures in program-controlled equipment (known as PCR machine) helps amplify DNA. The final product (known as amplicon) can be detected at the end through agarose gel electrophoresis or analyzed through different detection methods on a real-time basis. PCR has a broad range of applications, including basic research, medical diagnostics, forensics, and agriculture.]

Published: Mar 30, 2022

Keywords: PCR; DNA; cDNA; Primers; Annealing; Taq polymerase; Extension

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