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The protoplast generation and transformation of T. reesei was performed as described previously [54]. Typically, 10 µg of linearized plasmid DNA (in 15-µl sterile ddH2O) was used for the transformation of 107 protoplasts (in 200 µl). Selection was performed as described previously [44]. Resulting candidates were subjected to homokaryon purification by streaking conidia on selection plates.
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