连续多重PCR牛胚胎性别鉴定

Sex determination of bovine embryo using consecutive and multiplex polymerase chain reaction (PCR)

本实验以BOV97M序列和牛的1.715卫星DNA序列设计引物进行PCR扩增,其中前者为雄性特异扩增,后者为雌雄共同扩增,以进行牛的性别鉴定。PCR扩增共进行43个循环,其中前11个循环为雄性特异扩增,在随后的33个循环中,再加入内标引物,即进行多重PCR扩增,其灵敏度可达到10pg级(约3个细胞)。

The BOV97M and bovine 1.715 satelite sequences were selected to amplify male and bovine-specific DNA, respectively. In all 43 cycles were conducted, the first 11 PCR cycles were done with male-specific primer followed by an additional 33 cycles with bovine-specific primer namely Consecutive and Multiplex Polymerase Chain Reaction (PCR). This PCR method was applied successfully to use templates of 10pg (about 3 cells).