Keywords
SB buffer
optimization
basidiomycete
non-commercial protocol
and molecular identification
How to Cite
Abstract
The extraction of high-quality DNA is an essential step in genetic and molecular studies. In fungi, the ITS region of ribosomal DNA has been established as a reference marker for taxonomic identification, phylogeny, and metabarcoding applications. However, DNA extraction in basidiomycetes such as Ganoderma faces challenges due to the rigidity of the cell wall and the presence of secondary compounds that compromise DNA purity. In this study, a DNA extraction protocol for Ganoderma sp. was evaluated and optimized by replacing SDS with CTAB and adjusting NaCl concentration. Six samples were analyzed in two groups with different tissue amounts (25 and 50 mg), assessing DNA quality by agarose gel electrophoresis and its suitability through PCR amplification of the ITS2 region. Results showed welldefined, contaminant-free DNA bands in all samples, with no significant differences in DNA quality between groups, if buffer volume was properly adjusted. ITS2 amplification was successful in 100% of the cases, yielding fragments of approximately 900 bp, larger than previously reported in the literature. This discrepancy may be explained by intraspecific variation or differences in primer selection. These findings confirm the effectiveness of the optimized protocol and highlight the need for further studies to deepen the understanding of the genetic diversity of Ganoderma sp. and its biotechnological potential.
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