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Related Experiment Videos

DNA damage caused by common cytological fixatives

M P Douglas1, S O Rogers

  • 1Environmental and Forest Biology, State University of New York, College of Environmental Science and Forestry, 1 Forestry Drive, Syracuse, NY 13210, USA.

Mutation Research
|June 26, 1998
PubMed
Summary

Formaldehyde severely damages plant and fungal DNA, preventing amplification. Glutaraldehyde offers better DNA preservation for molecular studies, while other fixatives show varying DNA integrity.

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Area of Science:

  • Molecular Biology
  • Genetics
  • Biochemistry

Background:

  • Preserving DNA integrity is crucial for molecular analyses in plant and fungal research.
  • Cytological fixatives are commonly used to preserve tissue morphology but can impact DNA quality.
  • Understanding the effects of different fixatives on DNA is essential for selecting appropriate methods.

Purpose of the Study:

  • To evaluate the impact of various cytological fixatives and a storage buffer on DNA integrity and amplifiability in plant and fungal tissues.
  • To determine the optimal fixation method for DNA extraction and subsequent molecular analysis, such as polymerase chain reaction (PCR).

Main Methods:

  • Tissues from nine plant and fungal species were treated with seven fixatives (formaldehyde, FAA, glutaraldehyde, Lavdowsky's fluid) and one storage buffer (SED).

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  • DNA was extracted from treated and untreated tissues, and its molecular weight was assessed.
  • DNA amplifiability was tested using PCR targeting the nuclear rRNA small subunit gene, followed by sequencing.
  • DNA base damage rates were quantified for each treatment.

    • Main Results:

    • Formaldehyde treatments (3.7%) resulted in complete DNA damage, with no successful PCR amplification.
    • Glutaraldehyde treatments (1%) yielded DNA with an average length of 20 kb and minimal base damage (<0.1%).
    • FAA at pH 3.0 caused significant base damage (7.6%–15.6%), while FAA at pH 7.0, Lavdowsky's fluid, and SED showed lower damage rates (0.0%–3.6%).
    • Untreated and SED-treated tissues yielded high molecular weight DNA (>50 kb).

    Conclusions:

    • Formaldehyde is unsuitable for DNA preservation in plant and fungal tissues intended for molecular analysis.
    • Glutaraldehyde is a superior fixative for maintaining DNA integrity and amplifiability compared to formaldehyde and FAA.
    • Fixative choice and pH critically influence DNA quality, necessitating careful consideration for downstream molecular applications.