Abstract: The aim of this study was to optimize a method for rapid extraction of fungal genome for polymerase chain reaction (PCR). The optimal conditions for the preparation of fungal PCR templates were determined by the optimization of NaOH lysis conditions, bacterial conditions and generalizability analysis: the results presented the optimal conditions for the preparation of fungal PCR templates were a diameter of 3.5 mm colonies lysed in 100 μL of 0.5 mol/L NaOH at 25 ℃ for 15 min on a static basis. The purity （OD260/OD280 ） and the mass concentration of genome extracted by this method were 1.8－1.9 and 150－350 ng/μL, which were suitable for PCR verification. The results of universality analysis showed that the success rate of PCR validation of different genes in a large number ofAspergillus nigertransformants was as high as 98.6%, this method was also applicable to the validation of genes in a variety of fungi, such asSaccharomyces cerevisiae,Aspergillus oryzaeandTrichoderma reesei, which has a great universal applicability. This method is simple and safe, and can effectively improve the efficiency of fungal gene editing.

Key words: genome extraction, filamentous fungi, alkaline lysis, polymerase chain reaction, universality