Abstract: To explore an efficient, stable, low-cost DNA extraction method fromRondotia menciana Moore, genomic DNA was extracted using CTAB, protein enzyme K and salting- out method, respectively. The extracted genomic DNA samples of three methods were tested using photometric analysis and COI gene amplification. The results showed that, all the three methods can be used for the total DNA extraction, which are suitable for the PCR amplification, but genome DNA extracted by CTAB method and protein enzyme K method were higher in purity and production, degraded slightly. So we should select the proper method according to the objective and condition in the study.