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Updated: Jul 13, 2026
Collection and Extraction of Occupational Air Samples for Analysis of Fungal DNA
Published on: May 2, 2018
Rapid isolation of fungal genomic DNA suitable for long distance PCR
S L De Maeseneire1, I N Van Bogaert, T Dauvrin
1Laboratory of Industrial Microbiology and Biocatalysis, Department of Biochemical and Microbial Technology, Ghent University, Coupure links 653, Gent, 9000, Belgium. Sofie.DeMaeseneire@UGent.be
Abstract:
A quick and reliable method for screening fungal transformants for specific genetic modifications is essential for many molecular applications. We have compared the applicability of a few rapid DNA extraction methods for Myrothecium and Aspergillus and tested the resulting DNA as to its suitability for PCR. For Myrothecium gramineum, the highest DNA concentration was obtained with the procedure described by N. Vanittanakom et al. (J Clin Microbiol 2002, 40: 1739-1742). For A. nidulans, concentrations higher than 100 ng/mul were reached with the glass bead, the LiCl, the boiling, the liquid N(2) and the protoplast-based method. Samples of M. gramineum resulting from the boiling and the liquid N(2) procedure were suitable for the amplification of fragments up to 2.3 kb. The direct use of mycelium from M. gramineum in the PCR tube can be employed for the reproducible amplification of fragments up to 1 kb. Amplification of fragments up to 4.3 kb requires the use of the Elongase Mix on samples extracted with the liquid N(2) procedure.
Insights
This study compares rapid DNA extraction methods for fungal genetic screening. The Vanittanakom method yielded the most DNA for Myrothecium gramineum, enabling PCR amplification.
Area of Science:
- Molecular Biology
- Mycology
Background:
- Efficient DNA extraction is crucial for screening fungal transformants.
- Polymerase Chain Reaction (PCR) requires high-quality DNA templates.
Purpose of the Study:
- To evaluate and compare rapid DNA extraction methods for Myrothecium and Aspergillus species.
- To assess the suitability of extracted DNA for PCR amplification.
Main Methods:
- Comparison of multiple rapid DNA extraction techniques including boiling, liquid nitrogen, glass bead, LiCl, and protoplast-based methods.
- DNA yield and concentration were quantified.
- PCR amplification of various DNA fragment sizes was performed.
Main Results:
- The Vanittanakom et al. method provided the highest DNA yield for Myrothecium gramineum.
- For Aspergillus nidulans, glass bead, LiCl, boiling, liquid N(2), and protoplast methods yielded DNA concentrations over 100 ng/µL.
- DNA from boiling and liquid N(2) methods supported amplification of up to 2.3 kb fragments in M. gramineum.
- Direct mycelium use allowed amplification of up to 1 kb fragments.
- Elongase Mix with liquid N(2) extracted DNA enabled amplification of up to 4.3 kb fragments.
Conclusions:
- Rapid DNA extraction methods vary in efficiency for different fungal species.
- Specific extraction protocols and PCR reagents are necessary for amplifying larger DNA fragments.
