鉴定牛早期胚胎性别PCR反应体系的建立及优化研究

Study on Sex Determination in Bovine Early Embryos by Multi-PCR

根据牛Y染色体特异重复序列设计了一对引物,同时设计了一对牛常染色体引物作内标引物,研究了引物浓度、dNTP浓度、M g2+浓度、酶浓度、退火温度以及循环次数等因素对PCR方法鉴定牛胚胎性别的影响。试验结果表明,引物浓度、退火温度和循环次数对该反应体系影响较大,而dNTP浓度、T aq酶含量以及M g2+浓度的影响较小。

A pair of bovine male-specific PCR primers were designed based on the sequence of bovine Y-specific repeat sequence and a pair of primers for a sequence common to both sexs were designed as internal control primers. Multi-PCR conditions were optimized by varying the following parameters individually and in combination：the concentration of primers, the concentration of dNTP, the concentrationof Mg^2＋, the content of Taq, the annealing temperature and the number of cycles in the program. The results indicated that the concentration of primers, the annealing temperature and the number of cycles have more effect on the efficiency of multi-PCR, but the concentration of dNTP,the concentration of Mg^2＋ and the content of Taq have fewer.