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United States Department of Agriculture

Agricultural Research Service

Research Project: SYSTEMATICS OF LEPIDOPTERA: INVASIVE SPECIES, PESTS, AND BIOLOGICAL CONTROL AGENTS

Location: Systematic Entomology

Title: Room temperature DNA storage with slide-mounted Aphid specimens

Authors
item Miller, Gary
item Carmichael, Andrew
item Farvet, C.
item Scheffer, Sonja

Submitted to: Insect Conservation and Diversity
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: November 2, 2011
Publication Date: May 8, 2012
Repository URL: http://onlinelibrary.wiley.com/doi/10.1111/j.1752-4598.2012.00207.x/pdf
Citation: Miller, G.L., Carmichael, A., Farvet, C., Scheffer, S.J. 2012. Room temperature DNA storage with slide-mounted Aphid specimens. Insect Conservation and Diversity. 6(3):447-451.

Interpretive Summary: Aphids are small, soft-bodied insects that cause millions of dollars of losses to agricultural crops annually through direct feeding and transmission of plant diseases. Conventional means of study requires preparation of microscope slides with embedded aphids that are examined under high-powered microscopes. Aphid DNA is also studied to determine species identities and their relationships. Extraction of aphid DNA often involves the destruction of the specimen and its DNA is subsequently stored in low temperature freezers. This research uses a nondestructive DNA extraction technique that maintains an intact voucher specimen and its associated DNA is stored at room temperature on filter paper along with the slide-mounted specimen. This new technique adds greater versatility to microscope slide collections and can be applied to many systems that require DNA storage. The findings are especially important to museum curators, Federal quarantine specialists, state entomologists, extension personnel, and scientists studying aphids.

Technical Abstract: Most of the conventional molecular studies of aphids destroy the specimen in order to extract DNA. This DNA is subsequently stored in low temperature freezers. Room temperature storage of DNA with microscope slide-mounted voucher material is demonstrated by developing a system that uses filter paper in conjunction with the nondestructively cleared voucher specimen. Both COI (~600bp region) and COII (~300 bp region) of COII were successfully amplified after each filter paper re-extraction (one week, 90 days, and one year from application to the paper) after the DNA was stored at room temperature. The filter paper storage system shows promise as a way to augment conventional microscope slide collections and has application in other biological collections. Furthermore, the system does not require low-temperature storage systems thus eliminating the probably of system failure associated with refrigeration. This research adds a new dimension to microscope slide-based collections andit can be applied to many systems that require DNA storage. Furthermore, this system shows great potential for international and local transport of DNA research material for subsequent study.

Last Modified: 12/21/2014