α,β-D-Constrained Nucleic Acids Are Strong Terminators of Thermostable DNA Polymerases in Polymerase Chain Reaction
2011

Strong Terminators for DNA Polymerases in PCR

publication Evidence: moderate

Author Information

Author(s): Martínez Olivier, Ecochard Vincent, Mahéo Sabrina, Gross Grégori, Bodin Pierre, Teissié Justin, Escudier Jean-Marc, Paquereau Laurent

Primary Institution: Institut de Pharmacologie et de Biologie Structurale, Unité Mixte de Recherche 5089, Centre National de la Recherche Scientifique, Toulouse, France

Hypothesis

Can α,β-D-Constrained Nucleic Acids (CNA) be used as strong terminators of polymerization in PCR?

Conclusion

CNA can effectively terminate the activity of proofreading DNA polymerases in PCR, while non-proofreading polymerases may bypass this modification.

Supporting Evidence

  • One CNA unit is skipped by DNA polymerases, while two CNA units stop proofreading polymerases.
  • Pfu DNA polymerase is completely blocked by two CNA units, while Taq DNA polymerase may occasionally synthesize full-length fragments.

Takeaway

This study shows that special building blocks called CNA can stop certain enzymes from making DNA, which could help scientists in their experiments.

Methodology

The study involved PCR experiments using different DNA polymerases and analyzing the results through gel electrophoresis and sequencing.

Limitations

The study did not provide statistical data on the proportions of different PCR products obtained.

Digital Object Identifier (DOI)

10.1371/journal.pone.0025510

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