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Rapid and efficient protocol for DNA extraction and molecular identification of the basidiomycete Crinipellis perniciosa
S.C.O. Melo, C. Pungartnik, J.C.M. Cascardo and M. Brendel
Departamento de Ciências Biológicas, Universidade Estadual de Santa Cruz, Ilhéus, BA, Brasil
Corresponding author: M. Brendel
Genet. Mol. Res. 5 (4): 851-855 (2006)
Received September 26, 2006
Accepted November 6, 2006
Published December 14, 2006

ABSTRACT. DNA isolation from some fungal organisms is difficult because they have cell walls or capsules that are relatively unsusceptible to lysis. Beginning with a yeast Saccharomyces cerevisiae genomic DNA isolation method, we developed a 30-min DNA isolation protocol for filamentous fungi by combining cell wall digestion with cell disruption by glass beads. High-quality DNA was isolated with good yield from the hyphae of Crinipellis perniciosa, which causes witches’ broom disease in cacao, from three other filamentous fungi, Lentinus edodes, Agaricus blazei, Trichoderma stromaticum, and from the yeast S. cerevisiae. Genomic DNA was suitable for PCR of specific actin primers of C. perniciosa, allowing it to be differentiated from fungal contaminants, including its natural competitor, T. stromaticum.

Key words: Genomic DNA extraction, Crinipellis perniciosa, PCR, Filamentous fungi


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