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

Research output: Contribution to journalArticle

23 Citations (Scopus)

Abstract

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.

Original languageEnglish
Pages (from-to)421-430
Number of pages10
JournalZoological Studies
Volume41
Issue number4
Publication statusPublished - 2002 Oct 1

Fingerprint

Anguillidae
restriction fragment length polymorphism
mitochondrial DNA
polymerase chain reaction
alleles
cytochrome b
eel culture
DNA
Anguilla (eels)
prediction
genes
eel
digestion
nucleotides
methodology
taxonomy
mutation
enzymes

Keywords

  • Anguilla
  • Elver
  • Identification
  • PCR
  • RFLP

ASJC Scopus subject areas

  • Animal Science and Zoology

Cite this

Molecular techniques to identify freshwater eels : RFLP analyses of PCR-amplified DNA fragments and allele-specific PCR from mitochondrial DNA. / Lin, Yeong Shin; Poh, Yu Ping; Lin, Si-Min; Tzeng, Chyng Shyan.

In: Zoological Studies, Vol. 41, No. 4, 01.10.2002, p. 421-430.

Research output: Contribution to journalArticle

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