Laboratories that perform PCR on a routine basis need to count on reliable DNA isolation methods. In locations where supply of DNA extraction kits depends on importation, having an in-house protocol is desirable. This is also important for laboratories limited by budget constraints. We present a low-cost DNA isolation protocol that incorporates well-known techniques, but that we have adapted to various animal tissues. We tested this protocol on animal blood and muscle, and on cell suspension from skin swabs. The results were comparable, in terms of amount and quality of DNA, to those obtained with two other commercially available methods. DNA retrieved with this protocol has been successfully employed for Sanger sequencing of gene PCR products from animal tissues and blood, as well as for PCR-based diagnosis of chytrid fungus in amphibians and blood parasites in birds.