Evaluation of specimen preservatives for DNA analyses of bees

Large-scale insect collecting efforts that are facilitated by the use of pan traps result in large numbers of specimens being collected. Storage of these specimens can be problematic if space and equipment are limited. In this study, we investigated the effects of various preservatives (alcohol solutions and DMSO) on the amount and quality of DNA extracted from bees (specifically Halictidae, Apidae, and Andrenidae). In addition, we examined the amount and quality of DNA obtained from bee specimens killed and stored at -80 degrees C and from specimens stored for up to 24 years in ethanol. DNA quality was measured in terms of how well it could be PCR-amplified using a set of mitochondrial primers that are commonly used in insect molecular systematics. Overall the best methods of preservation were ultra-cold freezing and dimethyl sulfoxide, but these are both expensive and in the case of ultra-cold freezing, somewhat impractical for field entomologists. Additionally, dimethyl sulfoxide was shown to have adverse effects on morphological characters that are typically used for identification to the level of species. We therefore recommend that the best alternative is 95% ethanol, as it preserves bee specimens well for both morphological and molecular studies.