双重引物PCR鉴定APPSWE转基因小鼠

Optimized duplex PCR sets in the identification of APPSWE transgenic mice

本实验旨在探讨一种简单易行,能有效识别APPSWE转基因小鼠基因检测中假阴性结果的方法。在PCR体系中设计两对引物,一对为APP基因特异性引物,另一对为根据其内源性管家基因β-actin设置的参照引物。利用同一个PCR反应体系,对两对引物进行扩增。结果显示,双重引物PCR扩增的特异性片段与单引物扩增片段完全吻合,利用该法检出的转基因阳性率高于单引物PCR检出的转基因阳性率。本方法简单易行,能够有效识别以往利用单一PCR检测出现的假阴性结果,并且具有很好的特异性和灵敏性,值得在转基因小鼠鉴定实验中推广。

In order to identify the genotypes of APPSWE transgenic mice well, optimized duplex PCR assays were used in the study. Two pairs of primers, one for APPSWE gene and other for normal housekeeper β-actin gene, were applied in the same PCR reaction system. The resuh showed that duplex PCR amplified fragments were completely corresponded to the amplified bands produced by PCR with single primer pair. This method provided higher detection rate for positive trangenic mice than single primer PCR. The duplex PCR method is very effective to recognize the false negations detected by single PCR. This method is simple and sensitive with specificity, and is worth popularizing in the experiments for the identification of transgenic mice.