Abstract: In order to filter out a fast, efficient, inexpensive method to extract the Mycoplasma hyopneumoniae DNA tempiate. In this study we compared Tissue DNA Kit, double distilled boiled method and phenol-chloroform method to choose a best way to extract Mhp DNA. The results showed that the three methods were suitable for the extraction of genomic DNA of the bacterium, however, double distilled boiling method could not extracted genomic DNA from the tiusse, the Tissue DNA Kit could extract out of genomic DNA purely from the tissue, but got a small quantity of DNA each time, phenol-chloroform method could extract large DNA of the tissue, but complicated to operate, vulnerable to pollution, and with a little impurity bands. Acorrding to the different templates, select the appropriate DNA preparation method which is beneficial to make the detection accurated, meanwhile improve the detection efficiency, to provide protection for the early disease detection.