Development of a multiplex microsatellite assay for parentage assignment in orange?spotted groupers ( <scp> <i>Epinephelus coioides</i> </scp> )

Establishment of reliable pedigree information different lineages is crucial for selective breeding and stock enhancement programs. However, owing to the absence a assay parentage assignment, such lacking in orange-spotted grouper (Epinephelus coioides), which an economically important coral-reef fish. In this study, we developed two multiplex microsatellite polymerase chain reaction systems that consisted 12 markers (seven system A five B) assignment groupers. The theoretical exclusion probability per locus ranged from 0.079 0.436 when both parents were unknown (E-1P) 0.191 0.614 one parent was known (E-2P). combined values loci 98.8% 100% (E-2P), respectively. accuracy test mixed family (15 dams sires, 226 offspring) 100%. addition, wild populations (79 individuals) added as interferential offspring, only four misassigned stock. success rate 98.69%. We provide microsatellite-based groupers, will be useful programs species maybe other