Characterization of 22 polymorphic microsatellite markers for seven-band grouper Epinephelus septemfasciatus developed u
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Characterization of 22 polymorphic microsatellite markers for seven-band grouper Epinephelus septemfasciatus developed using a 454 pyrosequencing approach Hye Suck An • Jae Kwon Cho • Kyong Min Kim Maeng Hyun Son • Jeong In Myeong • Chul Min An
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Received: 27 February 2014 / Accepted: 24 March 2014 Ó Springer Science+Business Media Dordrecht 2014
Abstract Seven-band grouper, Epinephelus septemfasciatus, is of considerable economic value and a candidate species of aquaculture in Eastern Asia. It is necessary to monitor the stocks for the sustainable use and aquaculture of the species. Monitoring genetic variation among marine resources necessitates the development of genetic markers. Here, we report 22 polymorphic microsatellite markers with 2–16 alleles per locus. The mean observed and expected heterozygosity per locus was 0.53 and 0.57, respectively. Cross-amplification revealed variability in Epinephelus bruneus and Epinephelus akaara. The 22 markers developed can be a valuable tool for genetic conservation of this species. Keywords Epinephelus septemfasciatus Microsatellite loci Cross-species amplification In recent years, serranid species have been shown to be ideal aquaculture candidates because of their scarcity in the wild, high market value and rapid growth in temperate to tropical regions (Pierre et al. 2008). In Korea, seven-band grouper, Epinephelus septemfasciatus, is highly prized, and efforts to increase the harvest yield have been focused on Electronic supplementary material The online version of this article (doi:10.1007/s12686-014-0176-x) contains supplementary material, which is available to authorized users. H. S. An (&) C. M. An Biotechnology Research Division, National Fisheries Research and Development Institute, Pusan 619-705, Korea e-mail: [email protected] J. K. Cho K. M. Kim M. H. Son J. I. Myeong Aquaculture Management Division, Southwest Sea Fisheries Research Institute, National Fisheries Research and Development Institute, Yeosu 556-823, Korea
artificial seedling production since 2008. It is vital to understand genetic diversity and population structure of natural populations for management and preservation of exploited fish populations, which can be achieved by molecular genetic analysis (Ward 2006). Here, we report characteristics and primers for 22 novel polymorphic microsatellite markers isolated for E. septemfasciatus, which should be useful for future conservation genetics. Genomic DNA was extracted using TNES-Urea buffer method from fin tissue of one seven-band grouper captured off the southern coast around Geoje, Korea. A wholegenome shotgun library was generated from 2 lg of genomic DNA with the GS DNA Library Preparation Kit (Roche Applied Science). Sequencing was carried with only one-fourth of the run using a Genome Sequencer FLX Titanium instrument (Roche Applied Science). Of the 39,586 (14.7 %) unique sequences containing repeats, a total of 2,500 microsatellite sequences with a minimum of eight di-, tri- or tetra-nucleotide perfect r
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