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

Abstract

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.