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Simultaneous analysis of six microsatellite markers in Atlantic cod ( Gadus morhua): a novel multiplex assay system
Madjid Delghandi1, Atle Mortensen, Jon-Ivar Westgaard
1Department of Aquaculture, Norwegian Institute of Fisheries and Aquaculture Research (Fiskeriforskning), Muninbakken 9-13, Breivika, NO-9291 Tromsø, Norway. Madjid.Delghandi@fiskforsk.norut.no
Abstract:
A novel hexaplex assay system including Gmo8, Gmo19, Gmo35, Gmo37, Tch11, and Tch12 microsatellites from Atlantic cod, consisting of trinucleotide or tetranucleotide repeat units, is introduced. All 6 loci were coamplified in a single reaction employing dye-labeled primers. Alleles from these loci were sized using an internal standard by automated sample processing in an ABI 310 Genetic Analyser. Amplified alleles in profiles containing selected microsatellites were typed clearly, providing easily interpretable results. Sequencing data indicated that alleles at all loci consisted of simple repeat units. This may help minimize the likelihood of stuttering upon polymerase chain reaction amplification. The results suggest that the presented hexaplex assay system may be a useful tool in a selective breeding program in which genetic identification will allow different genotypes to be reared together from fertilization. This should have a great impact as it will make selective breeding more efficient.