Comparison of DNA Extraction Methods and Species Identification of Fish Meat
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Abstract:
In this study, an improved boiling method (BP method) and traditional proteinase K method (TP method) were used to extract DNAs from 16 kinds of fish meat samples. Then, the concentration and purity of the extracted DNAs as well as the performance of the common polymerase chain reaction (PCR) and fluorescence PCR were compared. In the mean time, the total DNA was extracted from 70 commercially available fish products using the BP method and analyzed for species identification using DNA barcode technology. The results showed that the extraction of DNA by the BP method required a shorter time. For most fish samples, the concentration of the DNA extracted by the BP method was lower than that by the TP method, but the two methods led to insignificant difference in the A260/A280 ratio of the extracted DNAs, thus, met the requirements for common and fluorescent PCR amplification. Moreover, the DNA extracted by the BP method could be used for DNA barcode identification of commercially available fish products. By comparison, 81.43% of the fish products could be matched with a similarity degree ≥ 98%, and the identification success rates at the species and genus levels were 48.57% and 72.86%, respectively. The BP method can shorten the DNA extraction time, reduce the extraction cost, and facilitate the detection of large quantities of samples.
