应用PCR法和LAMP法鉴定牛早期胚胎性别的研究

通过设计特异性引物，优化Mg^2＋浓度、退火温度和循环时间等影响PCR的因素，确定了适合现场应用的PCR反应体系；46枚胚胎现场的鉴定率为87％，移植妊娠率为28％，准确率为85％，证明建立的PCR胚胎性别鉴定方法可用于生产。使用LAMP法对126枚常规胚胎和42枚性控胚胎现场鉴定，鉴定率95％，移植妊娠率33％，准确率88％，结果证明该方法可应用于牛早期胚胎性别鉴定。PCR法和LAMP法所鉴定同一样品的结果一致，说明在控制好实验条件的情况下，根据实际情况可选用这两种方法中的任何一种进行性别鉴定应用。

Study on Sex Determination of Bovine Early Embryo by PCR and LAMP

By designing specific primer,optimizing Mg~(2 )concentration,annealing temperature and cycle condition,the optimal reaction system of PCR was established,which could be applied for the sex identification of bovine embryos in the field.The sex determining results of 46 embryos in the field were identification rate 87%,embryo transfer pregnancy rate was 28% and accuracy rate was 85%.It proved that the method of PCR could be applied in the field.Using LAMP method,126 normal embryos and 42 sex identified embryos in the field were determined.The identification rate was 95%,Embryo transfer pregnancy rate was 33% and accuracy rate was 88%.It proved that the method of LAMP could be applied for the bovine sex determination.The result of identifying the same sample by PCR and LAMP was consistent with each other.That means,any one of the two methods PCR and LAMP could be chosen to conduct sex determination according to the practical condition,if the lab condition was controlled quite well.