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Comparison of methods in the recovery and amplificability of DNA from fresh and processed sardine and anchovy muscle tissues
Authors:Nadia Besbes  Sami Fattouch  Saloua Sadok
Affiliation:1. Marine Biodiversity and Biotechnology Laboratory, Institut National des Sciences et Technologies de la Mer (INSTM), Tunisia;2. Laboratoire de Génie Biologique, Institut National des Sciences Appliqués et de Technologies (INSAT), Tunisia
Abstract:An important condition for a successful PCR amplification is an efficient DNA-extraction procedure out of a complex biological matrix such as canned fish. In this study we compared six extraction methods, including commercial kit, in terms of DNA yield, purity and time requirement. Such methods were applied to distinguish small pelagic fish species (Sardina pilchardus and Engraulisencrasicolus) among commercial canned products. The quantity and quality of DNA extracted were evaluated using the ratio A260/A280. Data were submitted to principal component analysis (PCA) in order to assess the differences between PCR results of fresh and processed anchovy and sardine muscles. Two main PC characterised the PCR of sardine and anchovy (70% and 69% of all variance): principal component 1 (PC1) (4% and 60%) and principal component 2 PC2 (66.0% and 9%) for sardine and anchovy, respectively. According to the PC1, the PCI/SDS and Chelex extractions (in decreasing order) were positively correlated with results of PCR for both species.
Keywords:Polymerase chain reaction   Canned fish   Sardina pilchardus   Engraulis encrasicolus   Mitochondrial DNA   Cytochrome b gene   Principal component analysis
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