AimsRecent molecular methods for diagnosis of superficial mycoses have determined the need for a rapid and easy method of extracting DNA. The aim of study was to determine growth conditions and techniques of DNA extraction for Microsporum canis, Trichophyton mentagrophytes and T. verrucosum. Methods and ResultsSamples were prepared of each of the DNA extraction methods (phenol-chloroform, CTAB and four different kits) for all of the incubation periods (4, 7 and 10days) of the cultures on the solid and in the liquid medium. The highest DNA concentrations were obtained using thephenol-chloroform method. The concentration of DNA extracted with the CTAB method accounted for 6221%, for kits it corresponded from 3553 to 1541%. The analysis of the DNA weight yield revealed the highest isolation efficiency of the phenol-chloroform method, 1mg of mycelium yielded 2238g DNA. Lower DNA yield (by 3932%) was obtained with the CTAB method; in the case of kits by 6846-8532%. In most of the techniques, the DNA yield on the solid medium was higher. ConclusionIn summary, the highest DNA yield was noted in the 7-day cultures and extraction with the phenol-chloroform method. Importantly, the type of culture was not relevant for the diagnostic result. Significance and Impact of the StudyMost mycoses are caused by fungi that reside in nature. The severity of the infection depends on the pathogenic attributes, socioeconomic factors and local environmental conditions. Recent diagnosis increasingly relies on not only the clinical features. Molecular identifications have determined the need for a rapid and easy method of extracting DNA. Usually two factors have to be considered: maximize the DNA yield and ensure that the extracted DNA is susceptible to enzymatic reactions. These data suggest that phenol-chloroform methods and a 7-day culture period may be useful for validation and constitute the first step of molecular diagnosis of dermatophytes.