Random Mutagenesis by Error-Prone Polymerase Chain Reaction Using a Heavy Water Solvent

Part of the Methods in Molecular Biology book series (MIMB, volume 1498)


Heavy water is a form of water that contains a heavier isotope of hydrogen (2H, also known as deuterium, D) or oxygen (17O or 18O). When using heavy water as a solvent, error-prone polymerase chain reaction (epPCR) can induce random mutations independent of the polymerase used or the composition of the PCR reaction mixture. This relatively new method can easily be combined with the existing epPCR methods to increase the rate of mutations.

Key words

Random mutagenesis Error-prone polymerase chain reaction (epPCR) Heavy water D2H218Stable isotope 


  1. 1.
    Kadonaga JT, Knowles JR (1985) A simple and efficient method for chemical mutagenesis of DNA. Nucleic Acid Res 13:1733–1745CrossRefPubMedPubMedCentralGoogle Scholar
  2. 2.
    Witkin EM (1976) Ultraviolet mutagenesis and inducible DNA repair in Escherichia coli. Microbiol Mol Biol Rev 40:869–907Google Scholar
  3. 3.
    Cadwell RC, Joyce GF (1994) Mutagenic PCR. Genome Res 3:S136–S140CrossRefGoogle Scholar
  4. 4.
    Biles BD, Connolly BA (2004) Low-fidelity Pyrococcus furiosus DNA polymerase mutants useful in error-prone PCR. Nucleic Acid Res 32:e176–e176CrossRefPubMedPubMedCentralGoogle Scholar
  5. 5.
    Minamoto T, Wada T, Shimizu I (2012) A new method for random mutagenesis by error-prone polymerase chain reaction using heavy water. J Biotechnol 157:71–74CrossRefPubMedGoogle Scholar

Copyright information

© Springer Science+Business Media New York 2017

Authors and Affiliations

  1. 1.Graduate School of Human Development and EnvironmentKobe UniversityKobeJapan

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