4组鱼类DNA条形码引物的筛选与优化

Optimization and selection for 4 fish DNA barcoding primer sets

目的通过对DNA条形码氧化酶亚基I基因(cytochrome oxidase subunit I,COI)的4组常用引物进行比较筛选,选择适于舟山鱼类鉴定的最佳引物。方法以舟山主产大黄鱼、小黄鱼、带鱼和乌贼样品DNA为模板,对4组常用DNA条形码引物进行PCR体系优化。通过比较PCR产物量、特异性、灵敏度和测序成功率,综合分析筛选最佳引物组。结果 4组引物均能扩增大黄鱼、小黄鱼和带鱼的DNA,对于乌贼DNA的扩增具有明显差异。COI优化引物具有易于扩增、测序简单等优势,更适于作为海产品鉴定的DNA条形码引物。结论本研究为舟山海产品的市场监管和实验室大量样品检测提供了技术参考。

Objective To select the best primers suitable for identification of fish in Zhoushan by comparing and screening 4 groups of commonly used primers of DNA barcode cytochrome oxidase subunit I（COI） gene. Methods The PCR system of 4 groups of common DNA barcode primers was optimized by taking the DNA samples of the main products of the fish, large yellow croaker, small yellow croaker, hairtail and cuttlefish from Zhoushan as the template. The optimal primer group was selected through comprehensive analysis by comparing the PCR product, specificity, sensitivity and sequencing success rate of PCR products. Results All 4 groups of primers could amplify the DNA of the large yellow croaker, the small yellow croaker and the hairtail, and the amplification of the cuttlefish DNA was significantly different. The COI optimized primer has the advantages of easy amplification and simple sequencing, which was more suitable for the identification of seafood DNA barcode primer. Conclusion This study provides a technical reference for the market supervision of Zhoushan marine products and the laboratory testing of a large number of samples.