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Updated: Jul 21, 2026

DNA-based Fish Species Identification Protocol
Published on: April 28, 2010
Comparison of different methods to produce single-strand DNA for identification of canned tuna by single-strand
H Rehbein1, I M Mackie, S Pryde
1Federal Research Center for Fisheries, Hamburg, Germany. 106616.766@compuserve.com
Abstract:
By using single-strand conformation polymorphism (SSCP) analysis of three amplicons of the cytochrome b gene obtained by the polymerase chain reaction (PCR) it was possible to differentiate between various species of tunas and bonitos processed as canned fish. Four different techniques were used to produce single-strand DNA (ssDNA): (i) Denaturation of double-strand DNA (dsDNA) by formamide and alkali, (ii) two-step asymmetrical PCR, (iii) one-step asymmetrical PCR, and (iv) exonuclease digestion of the phosphorylated strand of dsDNA. The technique rendering optimal results depended on the type of amplicon (i.e. the sequence).
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