Molecular - PCR - PCR reaction stages

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Molecular Pathology

Polymerase Chain Reaction

PCR reaction stages

Author: Rodney E. Shackelford, D.O., Ph.D. (see Reviewers page)

Revised: 22 September 2012, last major update July 2010

General

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● The thermocycles in PCR can be divided into three stages

● Understanding where each of these stages occurs in each PCR reaction is important, especially for the analysis of end-point PCR reactions

Exponential amplification:

● Initially, all the reagents employed (primers and nucleoside triphophates) in PCR are abundant and the Taq polymerase is fully functional

● Therefore the initial phases of PCR are characterized by near 100% efficient DNA amplification, with the amount of replicated DNA doubled per thermocycle

Leveling off stage:

● In later thermocycles the Taq polymerase begins to lose its activity and the concentration of primers and nucleoside triphophates become rate-limiting, hence the reaction rat significantly slows

Plateau:

● Eventually no new DNA amplification occurs due to exhaustion of the reagents and loss of enzyme activity

End of Molecular Pathology > Polymerase Chain Reaction > PCR reaction stages

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