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Title: LONG TERM PRESERVATION OF DNA FROM HONEY BEE (APIS MELLIFERA) COLLECTED IN AERIAL PITFALL TRAPS

Author
item Rubink, William
item MURRAY, K - UT PAN AMERICAN
item BAUM, KRISTEN - TEXAS A&M UNIVERSITY
item PINTO, M - TEXAS A&M UNIVERSITY

Submitted to: Texas Journal of Science
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: 1/2/2003
Publication Date: N/A
Citation: N/A

Interpretive Summary: A common method of collecting insects in many ecological studies involves the use of "pitfall" traps, placed in the soil where insects of interest may fall into them. Insects fall into a bath of alcohol, soapy water, polyethylene or propylene glycol where they are killed and temporarily preserved. New advances in molecular biology (e.g., the ability to study DNA in field-captured insect specimens) can provide much new information about insects collected in pitfall traps. In this study it was demonstrated that by using ecologically safe propylene glycol (commercially available as "antifreeze") both mitochondrial and nuclear genetic material are preserved adequately for analysis. Field and lab tests on honey bees collected and preserved in specially-designed aerial pitfall traps showed good preservation of the DNA after periods as long as 45 days and temperatures of 40 degrees C.

Technical Abstract: Newly developed molecular genetic analysis techniques can provide unique insights into the complex interrelationships among organisms, but they are dependent on collection and preservation techniques that adequately preserve DNA in the collected specimens. In this study we assessed the preservation of nuclear and mitochondrial DNA from honey bee (Apis mellifera L.) specimens which were first kept in propylene glycol-based antifreeze under various conditions, and then stored long-term, refrigerated in 95% ethanol. Two sets of bees were subjected to the propylene glycol treatment, then ethanol storage. One set consisted of bees captured in the field in propylene glycol-containing "scout traps" where they remained for up to 21 days. A second set consisted of bees taken from a hive and kept in propylene glycol under various temperature and lighting conditions then stored long-term in ethanol before evaluation of the persistence of nuclear and mitochondrial DNA using the polymerase chain reaction. DNA results demonstrate that propylene glycol-captured, ethanol-preserved honey bees retain both nuclear and mitochondrial DNA after capture and long term preservation. We suggest that with little or no modification the techniques described here might be applied to other studies involving trap-collected arthropod specimens.