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Five Atlantic cod ( Gadus morhua L.) microsatellites ( Gmo8, Gmo19, Gmo35, Gmo37 and Tch11) were selected for parentage assignment. All markers were co-amplified using dye-labelled primers and sized by automated sample processing in a 3130 xl Genetic Analyser. The genotyping of samples was reproducible and alleles from all loci could be genotyped confidently (precision and accuracy above 99.7 and 99.1% respectively). The number of alleles, observed allelic and genotypic frequencies were calculated. A population database was established for fish originating from randomly selected wild-caught Norwegian coastal- and North-East Arctic cod. Combined paternity exclusion and the probability of match were 99.97 and 1.5 × 10−9 respectively. The multiplex assay system was used successfully for parental assignment of 2336 juveniles from different mass spawning breeding tanks. Of all offspring, 91.2% were assigned to a single parental pair. In total, 523 different matings were recorded, and 69 of 70 dams and all of the 30 sires contributed to the offspring. [ABSTRACT FROM AUTHOR]