A simple and rapid DNA extraction protocol for filamentous fungi efficient for molecular studies

Abstract

A simple and rapid protocol for extracting high-quality DNA from filamentous fungi was studied. The method involved disruption of fungal cells by employing glass bead method, followed by inactivation of proteins using CTAB/proteinase K. The DNA yield from <span style="mso-bidi-font-style:italic">fungal isolates varied from 310-1879 µg g^-1 dry mycelium and a clear intact DNA band was observed upon agarose gel electrophoresis. Absorbency ratios (A₂₆₀/A₂₈₀) for DNA ranged 1.7-1.9, which indicated minimal presence of contaminating metabolites. PCR analysis like 18S <i>rRNA</i> gene amplification, random amplified polymorphic DNA (RAPD) and PCR-restriction fragment length polymorphism (PCR-RFLP) showed that DNA was compatible for downstream applications. This method can be applied to extract genomic DNA of filamentous fungi from different environmental sources.

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