Quantitative Fluorescent PCR (QF-PCR) on microsatellites, a fast and quantitative method to assay chimeric DNA samples in honeybees (Apis mellifera)Monica Bergema, Arne Rosethb, Sigbjørn Liena, b and Randi Aamodta
a Department of Animal and Aquacultural Sciences, Norwegian University of Life Sciences, 1432 Aas, Norway
b Center for Integrative Genetics (CIGENE), Norwegian University of Life Sciences, 1432 Aas, Norway
(Received 15 July 2005 - Revised 8 December 2005 - Accepted 22 December 2005 - published online 12 September 2006)
Abstract - The production of chimeras by nuclear transfer or cell transplantation requires functional tools for the detection of individuals that have successfully incorporated donor material. Quantitative Fluorescent PCR (QF-PCR) on microsatellites was employed on artificial chimeric honeybee DNA samples, to develop protocols for detection and quantification of donor genotypes after nuclear transfer and cell transplantations into honeybee embryos. Standard curves were produced for three microsatellite markers and demonstrate that there is a close to linear correlation between the amounts of donor genotypes estimated with QF-PCR and actual amounst of genotypes. The method was sensitive down to a 1% level and could detect and quantify small amounts of donor DNA in artificially mixed samples. QF-PCR offers fast and sensitive detection and quantification of genotypes and, thus, provides a useful tool for studies of chimeric honeybees and other species.
Key words: Quantitative Fluorescent PCR (QF-PCR) / microsatellites / nuclear transfer / honeybee / Apis mellifera
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© INRA, DIB-AGIB, EDP Sciences 2006