DNA extraction from the fungal organisms Coprinopsis Cinerea and Talaromyces Emersonii - Bertin Instruments

    Microbiology

    DNA extraction from the fungal organisms Coprinopsis Cinerea and Talaromyces Emersonii

    Sources: Institute of Technology, Sligo, Ireland

    Context

    DNA extraction is an essential tool in the molecular analysis of fungi. The architecture of the fungal cell wall renders many fungi resistant to standard DNA extraction procedures employed for yeast and bacteria. Fungal DNA extraction protocols tend to be time consuming and/or result in a poor yield of DNA. While commercial kits are available for the extraction of fungal DNA, they do require a pre step of grinding (with or without liquid N2) for the initial breaking up of mycelia. Grinding can be time consuming, taking up to 30 minutes per sample depending on the fungal species. For high throughput screening and analysis, a rapid DNA extraction method is required for fungal samples. This method was tested on two different classes of fungal species; an ascomycete (T. emersonii) and a basidiomycete (C. cinerea).

    Results

    DNA extractions were performed on both T. emersonii and C. cinerea mycelia harvested from liquid culture. The combination of the Minilys® personal homogenizer with the E.N.Z.A fungal DNA extraction kit resulted in a successful and rapid DNA extraction from both organisms. In order to increase efficiency for high throughput screening, the method was then tested on mycelia scraped from solid state cultures. DNA was successfully extracted from both organisms thus eliminating the need for liquid cultivation.

    Results

    References:
    Binninger, D. M., Skrzynia, C., Pukkila, P. J. and Casselton, L. A. (1987) DNA-mediated transformation of the basidiomycete Coprinus cinereus. Embo j, 6 (4), 835-40.
    Gupta, V.K., Tuohy, M.G., Ayyachamy, M., Turner, K.M. and O’Donovan, A.(2012) Laboratory Protocols in Fungal Biology: Current Methods in Fungal Biology. Springer Science & Business Media. p. 490-491.

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