ARS | Publication request: An economical method of indirect labeling of microsatellite primers

Submitted to: Journal of Apicultural Research
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: March 7, 2008
Publication Date: N/A

Interpretive Summary: The honey bee research community is expanding research in honey bee molecular biology, which can be a costly endeavor. Identification of quantitative trait loci (QTL) for traits of economic importance, to be used in marker-assisted breeding, is an important part of this research. The method of indirect labeling of microsatellite markers mentioned by Waldbieser et al. for catfish was modified and found to work for honey bees. The economical, high-throughput system for detecting microsatellites reported by Wang et al. was adopted for screening and detection. This system of indirect labeling and detection of microsatellites in a polyacrylamide gel provides an economical, flexible and relatively simple method of screening and genotyping with microsatellite markers.

Technical Abstract: In order to reduce the cost of labeling and detection of microsatellite markers for QTL research, an economical method of indirect labeling reported for catfish was modified to work with honey bees. The forward primer of each pair is modified by adding a 19-base sequence at the 5’ end. This same 19-base sequence is 5’ end labeled with Cy5 amidate and used as a universal primer in PCR reactions. Detection of PCR products is with an automated DNA sequencer for small allele differences and with an economical, high-throughput polyacrylamide gel for screening primers and detecting microsatellites with larger allele differences.