DNA extraction method for PCR in mycorrhizal fungi.
OBJECTIVE
To develop a simple and rapid DNA extraction protocol for PCR in mycorrhizal fungi.


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.


CONCLUSIONS
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.


CONCLUSIONS
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.
Manian
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Department of Plant Pathology and Microbiology, Horticulture Research International, Wellesbourne, Warwickshire CV35 9EF, UK.
Sreenivasaprasad
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Mills
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PR
D004271:DNA, Fungal / Q000235:genetics / Q000302:isolation & purification*; D005658:Fungi / Q000145:classification / Q000235:genetics* / Q000302:isolation & purification*; D016133:Polymerase Chain Reaction / Q000379:methods*
D004271:DNA, Fungal