Comparison of methods in the recovery and amplificability of DNA from fresh and processed sardine and anchovy muscle tissues

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. Statistical results confirmed that the quality of DNA, with the highest amplicon length obtained from the fresh and the different canned fish, was best preserved using the SDS/PCI precipitation method.