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

摘要

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 fungal isolates varied from 310-1879 mu g g(-1) dry mycelium and a clear intact DNA band was observed upon agarose gel electrophoresis. Absorbency ratios (A(260)/A(280)) for DNA ranged 1.7-1.9, which indicated minimal presence of contaminating metabolites. PCR analysis like 18S rRNA 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.