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Development of RAPD-SCAR and RAPD-generated PCRRFLP Markers for Identification of Four Anguilla eel Species  

Kim, Woo-Jin (Biotechnology Research Division, National Fisheries Research and Development Institute)
Kong, Hee-Jeong (Biotechnology Research Division, National Fisheries Research and Development Institute)
Kim, Young-Ok (Biotechnology Research Division, National Fisheries Research and Development Institute)
Nam, Bo-Hye (Biotechnology Research Division, National Fisheries Research and Development Institute)
Kim, Kyung-Kil (Biotechnology Research Division, National Fisheries Research and Development Institute)
Publication Information
Animal cells and systems / v.13, no.2, 2009 , pp. 179-186 More about this Journal
Abstract
Discriminating between eel species of the genus Anguilla using morphological characteristics can be problematic, particularly in the glass eel and elver stages. In this study, sequence-characterized amplified region (SCAR) and polymerase chain reaction (PCR)-restriction fragment length polymorphism (RFLP) markers were developed for the identification of Anguilla japoniea, Anguilla btcoior bicaor. Anguilla rostrata, and Anguilla anguilla. Random amplified polymorphic DNA (RAPD) fragments from A. japoniea (362 bp), A. bicolor bicctor (375 bp), A. rostrata (375 bp), and A. anguilla (375 bp) were isolated, sequenced, and converted to SCAR markers. The principal difference between the SCARs of A. japoniea and the three other species is the absence of a 13 bp deletion in the A. japoniea SCAR. Specific PCR primers amplified a 290 bp fragment for A. japoniea and 303 bp fragments for A. bicolor bicoior. A. rostrata, and A. anguilla. Restriction enzyme digestion with Taql, Mael, and Tru9l yielded PCR-RFLP patterns with differences that, when analyzed together, are sufficient for distinguishing each of the four eel species. In addition, RAPD fragments for A. japoniea (577 bp), A. bicoior bicoor (540 bp), A. rostrata (540 bp), and A. anguilla (509 bp) were also isolated and sequenced. The A. japoniea, A. bicoior blcoior. A. rostrata, and A. anguilla PCR products contain ten, nine, nine, and eight tandem repeats, respectively, of a 37 bp sequence. These results suggest that SCAR and PCR-RFLP markers and repeat numbers for specific loci will be useful for the identification of these four Anguilla eel species.
Keywords
Anguilla; SCAR; PCR-RFLP; species identification;
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