Comparison of DNA extraction methods in Ascomycota fungi

  • T. S. Shut Institute of Food Biotechnology and Genomics, Natl. Acad. Sci. Ukraine, Ukraine, 04123, Kyiv, Baidy-Vyshnevetskoho str., 2А https://orcid.org/0009-0008-3849-599X
  • I. A. Kosolapov Educational and Scientific Center "Institute of Biology and Medicine", Taras Shevchenko National University of Kyiv, Ukraine, 03022, Kyiv, Academika Hlushkova Ave., 2 https://orcid.org/0009-0004-7169-0894
DOI: https://doi.org/10.7124/FEEO.v37.1750
Keywords: Ascomycota, DNA extraction, SDS, CTAB, magnetic sorbent, spectrophotometry

Abstract

Aim. To compare DNA extraction methods from fungi of the Ascomycota. Protocols using SDS (sodium dodecyl sulfate), CTAB (cetyltrimethylammonium bromide), which were modified for this type of organisms, and the commercial kit NeoPrep¹⁰⁰ DNA Magnet_Plant (NEOGENE, Ukraine) were applied. Methods. Isolation of DNA from fungal mycelium using the three protocols mentioned above. Amplification of β-tubulin gene fragments using polymerase chain reaction with specific primers. Separation of amplicons in a 1.5 % agarose gel in the presence of ethidium bromide. Results. Various DNA extraction protocols were adapted for fungi of the Ascomycota. The concentration and quality of the extracted DNA were determined, which varied depending on the method used. It was established how different DNA extraction methods can affect polymerase chain reaction. Conclusions. It was found that the SDS method was the most effective and optimal for DNA isolation from fungi of the Ascomycota. The longest isolation time was the CTAB method, and the shortest was DNA isolation using the NeoPrep¹⁰⁰ DNA Magnet_Plant kit (NEOGENE, Ukraine).

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