Requirements for the application of protein sodium dodecyl sulfate-polyacrylamide gel electrophoresis and randomly amplified polymorphic DNA analyses to product speciation.
Raw, cooked, fried, smoked and gravad (brine-cured) products were analyzed by Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) of proteins and by randomly amplified polymorphic DNA (RAPD) in order to identify the species used in their manufacture. The discriminatory power of SDS-PAGE was dependent primarily on the composition and secondarily on the size of the gels: the Laemmli buffer system with 15% acrylamide and 0.087% piperazine diacrylamide separating gels resolved more discriminant protein bands than any of the commercial gels tested. Some of the processing conditions induced alterations in the protein patterns that made identification dubious. Differentiation even between closely related species was easier by RAPD than by SDS-PAGE. Neither the processing conditions nor the tissue from which the DNA was extracted had a significant effect on the RAPD profiles. For identifications based on SDS-PAGE, one should use an optimized gel composition and separate the sample under analysis in the same gel as the references. For RAPD-based identifications, the unknown sample should be amplified together with reference samples and separated in the same gel.
Martinez
I
I
Norwegian Institute of Fisheries and Aquaculture, Tromsø. iciar.martinez@fiskforsk.norut.no
Friis
T J
TJ
Seppola
M
M
D000818:Animals; D004247:DNA / Q000032:analysis* / Q000145:classification; D004591:Electrophoresis, Polyacrylamide Gel / Q000379:methods; D005511:Food Handling; D011506:Proteins / Q000032:analysis* / Q000145:classification; D019105:Random Amplified Polymorphic DNA Technique / Q000379:methods; D012967:Sodium Dodecyl Sulfate; D013045:Species Specificity
D011506:Proteins; D012967:Sodium Dodecyl Sulfate; D004247:DNA