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Non-destructive sources of DNA used to genotype honey bee (Apis mellifera) queens
Authors:Pamela G Gregory  & Thomas E Rinderer
Institution:United States Department of Agriculture, Agricultural Research Service, Honey bee Breeding, Genetics &Physiology Laboratory, 1157 Ben Hur Road, Baton Rouge, Louisiana 70820-5502, USA
Abstract:We describe a method for genotyping honey bee queens Apis mellifera L. (Hymenoptera: Apidae), using biological materials that are normally cast off during development (larval and pupal exuviae), or can be removed without apparent damage to queen longevity or acceptability to workers (wing clippings). Highly polymorphic microsatellite loci were successfully amplified from DNA from all of these sources, although with differing degrees of success. DNA was extracted using a simple Chelex 100® boiling procedure. Four microsatellite primers were used to amplify the DNA, and the PCR products were visualized on an ALFexpress Automated Sequencer. Genotypes created from these sources were consistent with those originating from tarsal tissue. Successful retrieval and amplification of DNA from the exuviae from immature queens allows potential breeding individuals to be genotyped and selected before they become adults. This procedure may therefore have value as DNA marker‐assisted breeding programs are developed for honey bees.
Keywords:Apis mellifera              breeding  Chelex  DNA extraction  genetic analysis  marker-assisted selection  PCR  Hymenoptera  Apidae
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