Multiplex PCR Detection of Exogenous Gene in Transgenic Mice

Focused on detection method for transgenic animals and its standardizations abroad in this article.It is essential to administrative regulations of transgenic animal and label of transgenic animal food.Extracting genomic DNA from mice tails by alkaline lysis method.The positive mice were screened by detecting of several transgenic elements such as cytomegalovirus(CMV) promoter,hGH polyA terminator,transmembrane protein 66(Tmem66)target gene with multiplex PCR method,as detecting mice endogenous GAPDH gene.The process of the multiplex PCR reaction was optimized,such as the optimal annealing temperature and the concentration of the PCR buffer,simultaneously studying the effect of reaction rate on multiplex PCR.The results showed that the multiplex PCR method was simple and accurate to detect transgenic elements of transgenic mice,which offered us useful experience for developing the typical testing standard of transgenic mammals and its drived food.