聚合酶链式反应-限制性片段长度多态性和芯片生物分析技术用于台湾海峡常见石斑鱼和鲷鱼的品种鉴定

Species identification of grouper and snapper in Taiwan Strait using polymerase chain reaction-restriction fragment length polymorphism analysis and lab-on-a-chip system

应用聚合酶链式反应-限制性片段长度多态性分析(PCR-RFLP)和芯片生物分析系统建立了台湾海峡常见石斑鱼和鲷鱼的分子生物学品种鉴定新方法。首先提取鱼的基因组脱氧核糖核酸(DNA)进行细胞色素b基因特定片段的PCR扩增,然后用DdeⅠ、HaeⅢ和NlaⅢ3种限制性内切酶进行酶切,在Agilent DNA1000芯片上对酶切片段进行分离。该方法成功鉴定了台湾海峡常见的8种石斑鱼品种和5种鲷鱼品种,是一种快速、简便、有效的鱼类品种鉴定分析手段。

Polymerase chain reaction-restriction fragment length polymorphism（PCR-RFLP） analysis and lab-on-a-chip system were used to identify grouper and snapper species in Taiwan Strait.A fragment of 464 bp length of mitochondrial cytochrome b gene was amplified by PCR and the products were digested with restriction enzymes DdeⅠ,HaeⅢ and NlaⅢ,individually.The fragments generated after digestion were further resolved on the DNA Chip.Eight grouper species and five snapper species were successfully identified.The results demonstrated that PCR-RFLP analysis and lab-on-a-chip system provide a fast,easy,automated,and reliable analysis approach.This approach is potential for the purpose of fish adulteration control.