Molecular techniques to identify freshwater eels: RFLP analyses of PCR-amplified DNA fragments and allele-specific PCR from mitochondrial DNA

Yeong Shin Lin, Yu Ping Poh, Si Min Lin, Chyng Shyan Tzeng

研究成果: 雜誌貢獻期刊論文同行評審

24 引文 斯高帕斯(Scopus)

摘要

Because of morphological ambiguity, traditional classification of freshwater eel elvers (of the genus Anguilla) has always been difficult and unreliable. This study analyzes 2 mitochondrial genes viz., cytochrome b and 12S ribosomal RNA (rRNA) genes from previous studies, to establish molecular standards for eel identification. Prediction of restriction fragment length polymorphism (RFLP) indicated that a combination of 3 enzyme results for cytochrome b revealed good resolution power. We also selected 39 specific nucleotide sites of the 2 genes at which the site is unique for individual species thus these can potentially be used for identification by polymerase chain reaction (PCR). To verify the accuracy of our predictions, we examined 58 specimens from 4 species by DpnII and HaeIII digestions and by PCR amplification using species-specific primers. Consistent with the putative mutation rates, more than 95% and 99% of the specimens could be successfully identified by the RFLP and PCR methods, respectively. The molecular techniques developed here can be helpful in eel aquaculture and ecological studies.

原文英語
頁(從 - 到)421-430
頁數10
期刊Zoological Studies
41
發行號4
出版狀態已發佈 - 2002 十月 1

ASJC Scopus subject areas

  • 動物科學與動物學

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