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United States Department of Agriculture

Agricultural Research Service

Research Project: Breeding, Genetics, Stock Improvement and Management of Russian Honey Bees for Mite and Small Hive Beetle Control and Pollination

Location: Honey Bee Breeding, Genetics, and Physiology Research

Title: An economical and effective high-throughput DNA extraction protocol for molecular marker analysis in honey bees

Authors
item Holloway, Beth
item Tarver, Matthew
item Rinderer, Thomas

Submitted to: Entomological Experimental Applied
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: May 7, 2013
Publication Date: September 1, 2013
Citation: Holloway, B.A., Tarver, M.R., Rinderer, T.E. 2013. An economical and effective high-throughput DNA extraction protocol for molecular marker analysis in honey bees. Entomological Experimental Applied. 148:196-200.

Interpretive Summary: We have developed a technique for extracting DNA from honey bee samples for use in analysis of honey bee genetics. This technique is extremely cost efficient, non-hazardous, and fast. It also requires much less labor in preparing the samples for the DNA extraction than traditional honey bee DNA extraction methods. By using a mixture of essentially salt, soap and rubbing alcohol, we can obtain high quality DNA that can be used directly for molecular biology analysis.

Technical Abstract: Extraction of DNA from tissue samples can be expensive both in time and monetary resources and can often require handling and disposal of hazardous chemicals. We have developed a high throughput protocol for extracting DNA from honey bees that is of a high enough quality and quantity to enable hundreds of PCR reactions from a single extract, yet is extremely cost effective and non-hazardous. By utilizing high sodium chloride concentrations we can decant contaminating proteins in a buoyant conglomeration while retaining a highly pure DNA pellet. This method generates sufficient DNA to enable downstream applications such as molecular marker analysis useful for QTL studies and mapping in honey bees.

Last Modified: 4/17/2014
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