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Thirty-four microsatellite loci were isolated from three reef fish species; golden snapper  Lutjanus johnii , blackspotted croaker  Protonibea diacanthus  and grass emperor  Lethrinus laticaudis  using a next generation sequencing approach. Both IonTorrent single reads and Illumina MiSeq paired-end reads were used, with the latter demonstrating a higher quality of reads than the IonTorrent. From the 1–1.5 million raw reads per species, we successfully obtained 10–13 polymorphic loci for each species, which satisfied stringent design criteria. We developed multiplex panels for the amplification of the golden snapper and the blackspotted croaker loci, as well as post-amplification pooling panels for the grass emperor loci. The microsatellites characterized in this work were tested across three locations of northern Australia. The microsatellites we developed can detect population differentiation across northern Australia and may be used for genetic structure studies and stock identification.

peerj

Characterization, development and multiplexing of microsatellite markers in three commercially exploited reef fish and their application for stock identification