An efficient and safe method for the extraction of total DNA from shed frog skin

An efficient protocol for the isolation of high-quality DNA from tissues is essential to many aspects of molecular biology. Shed skin is often overlook as a source for high-quality DNA in molecular studies involving frog. In this study, DNA from shed skin of frog was extracted using phenol–chloroform, the DNeasy extraction kit (Qiagen) and an improved salting-out protocol, in comparison with those of skeletal muscle and toe. We found that the improved salting-out procedure obtained the highest DNA yield, and avoided using toxic chemicals such as phenol, chloroform, isoamyl alcohol, and/or guanidine isothiocyanate. The quantity of DNA from shed skin was lower compared to other tissue in Rana dybowskii . Nevertheless, DNA from shed skin was useful for amplifing mtDNA and nuclear DNA using PCR, as were Rana amurensis and Bufo gargarizans . These results indicate that extrating DNA from shed frog skin by improved salting-out protocol is an efficient and safe method for the conservation genetic studies and population genetics without destroy in frogs.