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Species Identification of the Tropical Abalone (Haliotis asinina, Haliotis ovina, and Haliotis varia) in Thailand Using RAPD and SCAR Markers

  • Klinbunga, Sirawut (Marine Biotechnology Research Unit, National Center for Genetic Engineering and Biotechnology (BIOTEC), National Science and Technology Development Agency (NSTDA)) ;
  • Amparyup, Piti (Program of Biotechnology, Faculty of Science, Chulalongkorn University) ;
  • Leelatanawit, Rungnapa (Program of Biotechnology, Faculty of Science, Chulalongkorn University) ;
  • Tassanakajon, Anchalee (Department of Biochemistry, Faculty of Science, Chulalongkorn University) ;
  • Hirono, Ikuo (Laboratory of Genome Science, Graduate School of Marine Science and Technology, Tokyo University of Marine Science and Technology) ;
  • Aoki, Takashi (Laboratory of Genome Science, Graduate School of Marine Science and Technology, Tokyo University of Marine Science and Technology) ;
  • Jarayabhand, Padermsak (Aquatic Resources Research Institute, Chulalongkorn University) ;
  • Menasveta, Piamsak (Marine Biotechnology Research Unit, National Center for Genetic, Engineering and Biotechnology (BIOTEC), National Science and Technology Development Agency (NSTDA))
  • Published : 2004.03.31
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Abstract

A randomly amplified polymorphic DNA (RAPD) analysis was used to identify the species- and population-specific markers of abalone; Haliotis asinina, H. ovina, and H. varia in Thai waters. Fifteen species-specific and six population-specific RAPD markers were identified. In addition, an 1650 bp band (UBC195) that was restricted to H. ovina from the Gulf of Thailand (east) was also found. All of the specific RAPD markers were cloned and sequenced. Twenty pairs of primers were designed and specificity-tested (N = 12 and 4 for target and non-target species, respectively). Seven primer pairs (CUHA1, 2, 4, 11, 12, 13, and 14) were specifically amplified by H. asinina DNA, whereas a single pair of primers showed specificity with H. ovina (CUHO3) and H. varia (CUHV1), respectively. Four primer pairs, including CUHA2, CUHA12, CUHO3, and CUHV1, were further examined against 216 individuals of abalone (N = 111, 73, and 32, respectively). Results indicated the species-specific nature of all of them, except CUHO3, with the sensitivity of detection of 100 pg and 20 pg of the target DNA template for CUHA2 and CUHA12 and CUHV1, respectively. The species-origin of the frozen, ethanol-preserved, dried, and boiled H. asinina specimens could also be successfully identified by CUHA2.

Keywords

References

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