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Updated: Sep 16, 2025

Development of a Quantitative Recombinase Polymerase Amplification Assay with an Internal Positive Control
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How reliable is PCR? A mini review.

Helen Konrad1, Andreas Beyer2

  • 1Institut für Rechtsmedizin, https://ror.org/04mz5ra38Universitätsklinikum Essen, Essen, Germany.

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PubMed
Summary

Polymerase Chain Reaction (PCR) is reliable for scientific research. Despite concerns about DNA thermolability during the denaturation phase, thermal hydrolysis does not impact PCR performance, specificity, or results.

Keywords:
DNA analysisPCRpseudoscientific critiquereliability of PCRthermostability of DNA

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Area of Science:

  • Molecular Biology
  • Biochemistry
  • Genetics

Background:

  • Early studies (1950s-60s) indicated DNA's thermolability.
  • This raised questions about DNA stability during Polymerase Chain Reaction (PCR) denaturation.
  • Potential DNA degradation could affect PCR reliability.

Purpose of the Study:

  • To investigate the impact of DNA thermolability on PCR reliability.
  • To determine if thermal hydrolysis affects PCR performance and specificity.
  • To validate the accuracy of contemporary PCR protocols.

Main Methods:

  • Review of historical physicochemical studies on DNA.
  • Analysis of thermal hydrolysis effects on DNA during PCR denaturation.
  • Presentation of primary data to demonstrate PCR reliability.

Main Results:

  • Thermal hydrolysis losses are negligible in modern PCR protocols.
  • DNA thermolability does not compromise PCR performance or specificity.
  • Primary data confirms the reliability of PCR results.

Conclusions:

  • Contemporary PCR protocols are robust and reliable.
  • Concerns regarding DNA thermolability in PCR are unfounded.
  • PCR remains a valid and significant tool in molecular biology.