TECHNICAL NOTE Isolation and characterization of fifteen novel microsatellite loci in golden mandarin fish (Siniperca scherzeri) Steindachne Min Yang • Xu-Fang Liang • ChangXu Tian • Yasmeen Gul • YaQi Dou • Liang Cao • Rui Yu Received: 15 November 2011 / Accepted: 2 January 2012 / Published online: 24 January 2012 Ó Springer Science+Business Media B.V. 2012 Abstract We described the isolation and characterization of 15 new microsatellite loci from golden mandarin fish, Siniperca scherzeri. The analysis of variability was per- formed in 34 individuals. Allelic diversity ranged from 2 to 7 alleles per locus, with observed and expected heterozy- gosities ranging from 0.294 to 1.000 and 0.261 to 0.740, respectively. Seven loci deviated significantly from Hardy– Weinberg equilibrium after Bonferroni correction, and no significant linkage disequilibrium were found among 15 pairs of loci following the Bonferroni correction. These are the first microsatellite markers characterized from the S. scherzeri. We expect these microsatellite markers to be useful for population genetic studies in this species. Keywords Siniperca scherzeri  Golden mandarin fish  Microsatellite loci  Genetic diversity Golden mandarin fish (Siniperca scherzeri), belongs to a group of the lower percoid fishes, is mainly distributed in the inland water of China (Li 1991; Zhou et al. 1988). Recently, the population of S. scherzeri has diminished due to over-fishing and greater anthropogenic interference in their habitat (Liang 1996) which might lead to the degen- eration of the genetic resources of the wild population. Therefore, the rational use of natural resources is urgently required in order to sustain the quality of broodstock. Microsatellites are short tandem repeated DNA sequences with a length of 1-6 bp (Weber and May 1989). Due to high polymorphism, even distribution on genomes, ease of genotyping by using PCR and co-dominance, microsatellites have been the marker of choice for a number of studies including genome mapping, genetic diversity and forensic studies (Goldstein and Schlotterer 1999). Micro- satellite loci of some economically important fish species such as Salminus hilarii (Silva and Hilsdorf 2011), gilthead seabream (Parati et al. 2011), turbot (Gu et al. 2009), Japanese Spanish mackerel (Xing et al. 2009) and whisker sheatfish (Phongkaew et al. 2011) have been isolated and characterized earlier. However, microsatellite loci from S. scherzeri have not yet been isolated. Microsatellites were isolated using a hybridization- based capture methodology, following the protocol described by Billotte et al. (1999). Briefly, genomic DNA was extracted from fin clips using the standard phenol/ chloroform method (Sambrook and Russel 2002) and digested with the restriction enzyme MseI (BioLabs). DNA fragments of 300-1000 bp were size selected by agarose gel electrophoresis and excised gel was purified by using PBZ0202-1 DNA purification kit (New probe biotech). Then specific adapters (5 0 -GACGATGAGTCCTGAG-3 0 and 5 0 -TACTCAGGACTCAT-3 0 ) were ligated to the digested DNA. Approximately amplified DNA fragments were hybridized with 5 0 - biotin-labeled oligonucleotide (CA) 14 probe and the streptavidin magnetic beads (Bio Lab) were used to capture the target fragments. The cap- tured DNA fragments were eluted from the beads-probe DNA mixture by treating it with TE buffer at 95°C for 5 min. The enriched DNAs were cloned into the pGEM-T plasmid vector (Promega, USA) and were transformed into competent Escherichia coli strain DH-5a (Promega, USA). A total of 112 positive colonies were identified and incu- bated overnight using LB broth and ampilillin, and then amplified. The presence of insert was verified by direct M. Yang  X.-F. Liang (&)  C. Tian  Y. Gul  Y. Dou  L. Cao  R. Yu College of Fisheries, Huazhong Agricultural University, Wuhan 430070, China e-mail: xfliang@mail.hzau.edu.cn 123 Conservation Genet Resour (2012) 4:599–601 DOI 10.1007/s12686-012-9601-1