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Comparison of different methods to produce single-strand DNA for identification of canned tuna by single-strand conformation polymorphism analysis.

Authors
  • Rehbein, H1
  • Mackie, I M
  • Pryde, S
  • Gonzales-Sotelo, C
  • Perez-Martin, R
  • Quinteiro, J
  • Rey-Mendez, M
Type
Published Article
Journal
Electrophoresis
Publication Date
June 1998
Volume
19
Issue
8-9
Pages
1381–1384
Identifiers
PMID: 9694285
Source
Medline
License
Unknown

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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