Objective To establish a simple，fast and economic total DNA extraction method to serve the rapid identification of model mouse genotype in large number of mice. Methods Four methods，i.e.alkali boiling routine method, alkali boiling simplified method, protease K lysis and DNA extraction kit methods were used to extract the total DNA from transgenic mice tails．The purity and concentrationof DNA obtained by the four methods were measured, the advantages and disadvantages of gel electrophoresis were evaluated, and the time-consuming and experimental costs of the four methods were compared．Results The DNA concentration extracted by protease K cleavage method is the highest, followed by alkaline boiling simplified method and conventional method; The DNA purity obtained from the reagent kit treatment is relatively high, followed by the alkaline boiling simplification method. The DNA templates obtained by four methods can be amplified by PCR and Gel electrophoresis to obtain clear DNA target bands, and can also be used for gene identification after being stored at -20 ℃ for one month. In addition, the simplified alkaline boiling method has the lowest cost and the least time consumption. Conclusion The alkaline boiling simplification method is currently the fastest and most economical DNA template extraction method for batch genotyping of model mice.