Introduction. The techniques presented allow a long-term conservation of Cylindrocladium strains to preserve them from major morphological and physiological changes. The principle of the methods applied, key advantages, starting plant material, time required and expected results are presented. Materials and methods. The necessary laboratory materials, and details of the nine steps achieved for the monospore purification of Cylindrocladium strains, their long-term maintenance, and the production of conidia and microsclerotia are described. Results. When revived, purified strains generate actively growing colonies within 48–72 h. Such colonies can then be readily used for conidia or microsclerotia production.