The fungal strains were grown as described previously (Libert et al. 2015 (link)). Briefly, the fungal strains were spiked into a S10 Sabouraud liquid medium (Biorad, Temse, Belgium) and were grown at 25 °C with constant agitation between 3 to 21 days according to the species’ growth conditions.
S10 sabouraud liquid medium
Manufactured by Bio-Rad
Sourced in Belgium
S10 Sabouraud liquid medium is a culture medium used for the isolation and cultivation of fungi. It provides nutrients and growth factors required for the growth of a wide range of fungal species.
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2 protocols using s10 sabouraud liquid medium
1
Fungal Strain Growth Conditions
2
Fungal DNA Extraction and Purification
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The fungal strains were grown in a S10 Sabouraud liquid medium (Bio-Rad, Temse, Belgium) at 25 °C with constant agitation between 3 and 10 days according to the species’ growth conditions.
After this incubation time, 300 mg of wet sample was transferred to cryotubes containing 0.25 ml of acid-washed glass beads (Sigma-Aldrich, Diegem, Belgium) put at −80 °C during 40 min and freeze-dried overnight with a freeze-dryer Epsilon 1-6D (Martin Christ, Osterode am Harz, Germany). Freeze-dried fungi were subsequently beat-beaten with a Mini bead beater (Biospec Products, OK, USA) during 1 min at maximal speed.
The total DNA was extracted with an adapted phenol chloroform (24:1) protocol (Ashktorab and Cohen 1992 (link)) and purified with the Qiagen CTAB genomic Tip-20 kit (Qiagen Benelux, B.V., KJ Venlo, the Netherlands) according to the manufacturer’s recommendation. DNA was eluted with 100 μl Gibco® DNase, RNase, protease free water (Life Technologies, Gent, Belgium). The DNA integrity was verified on a 2 % agarose gel. The DNA amount and purity were evaluated with a Nanodrop® 2000 (Thermo Scientific, Wilmington, USA).
After this incubation time, 300 mg of wet sample was transferred to cryotubes containing 0.25 ml of acid-washed glass beads (Sigma-Aldrich, Diegem, Belgium) put at −80 °C during 40 min and freeze-dried overnight with a freeze-dryer Epsilon 1-6D (Martin Christ, Osterode am Harz, Germany). Freeze-dried fungi were subsequently beat-beaten with a Mini bead beater (Biospec Products, OK, USA) during 1 min at maximal speed.
The total DNA was extracted with an adapted phenol chloroform (24:1) protocol (Ashktorab and Cohen 1992 (link)) and purified with the Qiagen CTAB genomic Tip-20 kit (Qiagen Benelux, B.V., KJ Venlo, the Netherlands) according to the manufacturer’s recommendation. DNA was eluted with 100 μl Gibco® DNase, RNase, protease free water (Life Technologies, Gent, Belgium). The DNA integrity was verified on a 2 % agarose gel. The DNA amount and purity were evaluated with a Nanodrop® 2000 (Thermo Scientific, Wilmington, USA).
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