Rapid methods for nucleic acids extraction from Petri dish-grown mycelia

Area of Science:

• Mycology
• Molecular Biology
• Biochemistry

Background:

• Filamentous fungi are crucial in various biological and industrial processes.
• Efficient isolation of nucleic acids (DNA and RNA) is essential for studying fungal genetics and function.
• Existing methods can be time-consuming, costly, or yield suboptimal results.

Purpose of the Study:

• To develop rapid, economic, and efficient methods for isolating nucleic acids from filamentous fungi.
• To optimize sample processing for high-throughput analysis.
• To assess the yield and purity of isolated DNA and RNA.

Main Methods:

• Direct recovery of mycelium from Petri-dish cultures.
• All experimental procedures conducted within microfuge tubes.
• High-throughput processing of up to 24 samples per day.
• Spectrophotometric quantification and gel electrophoresis for purity and yield assessment.

Main Results:

• Successful isolation of nucleic acids from filamentous fungi using a streamlined protocol.
• High yields of total DNA and RNA (up to 100 micrograms) with sufficient purity for downstream applications.
• Demonstrated efficient processing of multiple samples within a single day.
• Significant recovery of mitochondrial DNA, visualized by ethidium bromide-stained gels.

Conclusions:

• The developed methods offer a rapid, cost-effective, and efficient approach for nucleic acid isolation from filamentous fungi.
• The protocol's simplicity and high throughput make it suitable for large-scale fungal genetic studies.
• The recovery of substantial amounts of pure DNA, including mitochondrial DNA, facilitates diverse molecular analyses in mycology.