INTRODUCTIONMany different DNA isolation methods have been employed successfully in ants. Parameters such as the size and developmental stage of the specimen (egg, larvae, or adult) and the subsequent use of the DNA will mostly determine which method should be used. Ant body sizes range from minute (1-2 mm in length) to large (30 mm), and the volume of the initial digestion should be adjusted accordingly. Whereas workers usually have low concentrations of storage proteins and fat, queens and larvae can contain considerable amounts of these substances that can interfere with the subsequent use of the isolated DNA. Ants also have many glands in the head and abdomen, and the contents of these glands can also interfere with the successful application of polymerase chain reaction (PCR) or restriction digests of the isolated DNA. This protocol presents two DNA isolation methods that have worked reliably for a wide range of ant species: a “quick and dirty” technique using Chelex isolation, and a more elaborate, classical phenol:chloroform procedure.