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Updated: May 21, 2026
06:53
Isolation, Characterization, and Total DNA Extraction to Identify Endophytic Fungi in Mycoheterotrophic Plants
Published on: May 5, 2023
A rapid and inexpensive method for isolation of total DNA from Trichoderma spp (Hypocreaceae)
J C Vazquez-Angulo1, V Mendez-Trujillo, D González-Mendoza
1Instituto de Ciencias Agrícolas, Universidad Autónoma de Baja California, Baja California, México.
Genetics and Molecular Research : GMR
|June 2, 2012
Summary
We developed a simple DNA extraction method for filamentous fungi. This protocol yields high-quality genomic DNA suitable for PCR amplification, overcoming common challenges like complex cell walls and contaminants.
Area of Science:
- Molecular Biology
- Mycology
- Biochemistry
Background:
- Filamentous fungi possess complex cell walls and secondary metabolites that impede high-quality genomic DNA extraction.
- Standard DNA extraction protocols often require maceration and ethanol precipitation, which can be inefficient for fungal samples.
Purpose of the Study:
- To develop an optimized and simplified protocol for extracting high-quality genomic DNA from filamentous fungi.
- To ensure the extracted DNA is pure and suitable for downstream molecular applications like PCR amplification.
Main Methods:
- A modified sodium dodecyl sulfate/phenol protocol was employed, omitting liquid nitrogen maceration and final ethanol precipitation.
- Mycelium was directly recovered from the culture medium for processing.
Main Results:
- The isolated DNA exhibited A260/280 absorbance ratios of approximately 1.7-1.9, indicating high purity.
- A260/230 values exceeded 1.6, demonstrating negligible contamination by polysaccharides and other metabolites.
- The DNA quality was sufficient for molecular applications, confirmed by its suitability for PCR amplification.
Conclusions:
- The developed method provides a robust and efficient means for genomic DNA extraction from filamentous fungi.
- This protocol is advantageous due to its simplicity, cost-effectiveness, and applicability to organisms with similar extraction challenges.
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