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Comparison of different methods to produce single‐strand DNA for identification of canned tuna by single‐strand conformation polymorphism analysis
Author(s) -
Rehbein Hartmut,
Mackie Ian M.,
Pryde Susan,
GonzalesSotelo Carmen,
PerezMartin Ricardo,
Quinteiro Javier,
ReyMendez Manuel
Publication year - 1998
Publication title -
electrophoresis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.666
H-Index - 158
eISSN - 1522-2683
pISSN - 0173-0835
DOI - 10.1002/elps.1150190830
Subject(s) - single strand conformation polymorphism , single strand , d loop , amplicon , microbiology and biotechnology , dna , biology , coding strand , polymerase chain reaction , transcription bubble , chemistry , polymerase , genetics , mitochondrial dna , gene , rna dependent rna polymerase
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).