Related Experiment Video
Updated: Jul 26, 2026
Collection and Extraction of Occupational Air Samples for Analysis of Fungal DNA
Published on: May 2, 2018
DNA extraction method for PCR in mycorrhizal fungi
S Manian1, S Sreenivasaprasad, P R Mills
1Department of Plant Pathology and Microbiology, Horticulture Research International, Wellesbourne, Warwickshire CV35 9EF, UK.
Aims:
To develop a simple and rapid DNA extraction protocol for PCR in mycorrhizal fungi.
Methods And Results:
The protocol combines the application of rapid freezing and boiling cycles and passage of the extracts through DNA purification columns. PCR amplifiable DNA was obtained from a number of endo- and ecto-mycorrhizal fungi using minute quantities of spores and mycelium, respectively.
Conclusion:
DNA extracted following the method, was used to successfully amplify regions of interest from high as well as low copy number genes. The amplicons were suitable for further downstream applications such as sequencing and PCR-RFLPs.
Significance And Impact Of The Study:
The protocol described is simple, short and facilitates rapid isolation of PCR amplifiable genomic DNA from a large number of fungal isolates in a single day. The method requires only minute quantities of starting material and is suitable for mycorrhizal fungi as well as a range of other fungi.
Insights
This study presents a fast DNA extraction method for mycorrhizal fungi, enabling quick PCR amplification. The protocol yields high-quality DNA suitable for various downstream applications, even from small samples.
Area of Science:
- Mycology
- Molecular Biology
- Genetics
Background:
- Mycorrhizal fungi are crucial for plant health.
- Efficient DNA extraction is vital for molecular studies of fungi.
- Existing protocols can be time-consuming and require significant starting material.
Purpose of the Study:
- To develop a simple, rapid DNA extraction protocol specifically for mycorrhizal fungi.
- To enable Polymerase Chain Reaction (PCR) amplification of fungal DNA.
Main Methods:
- The protocol utilizes rapid freezing and boiling cycles.
- DNA purification columns are employed for isolating amplifiable DNA.
- The method requires only minute quantities of fungal spores or mycelium.
Main Results:
- PCR-amplifiable DNA was successfully extracted from various endo- and ecto-mycorrhizal fungi.
- The extracted DNA yielded successful amplification of high and low copy number genes.
- Amplicons were suitable for downstream applications like sequencing and PCR-Restriction Fragment Length Polymorphisms (PCR-RFLPs).
Conclusions:
- The developed protocol is simple, rapid, and efficient for isolating PCR-amplifiable genomic DNA.
- It allows processing of numerous fungal isolates within a single day.
- The method is versatile, applicable to mycorrhizal fungi and other fungal species.
