Insect DNA Extraction Protocol

Abstract

The following procedure for obtaining insect DNA is similar to a plant DNA extraction procedure (Saghai-Maroof et al. 1984) and relies on a nonionic detergent, hexadecyltrimethylammonium bromide (CTAB) to lyse cells. The resulting DNA is suitable for use as a template in polymerase chain reactions to generate RAPD markers or for Southern blotting. This general protocol has been successfully applied for RAPD analyses of bees (Apis, Bombus, Frieseomelitta, Trigona, Scaptotrigona, Melipona), wasps (Vespid species) ants (Leptothoraxy Pogonomyrmex), beetles (Tribolium) and mites (Varroa). The method used to extract honey bee DNA is presented here. Modifications in grinding techniques may be necessary for different species of arthropods. Grinding in a mortar and pestle with liquid nitrogen will usually improve yields and produce higher molecular weight DNA for Southern blotting, but the simple grinding method described here is adequate to provide template for PCR.