Authors : Huabin Zhu, Bo Lin, Jun Chen, Haisheng Hao, Xueming Zhao, Shujing Li, Weihua Du, Tong Qin, Yan Liu and Dong Wang
Abstract: A rapid and simple PCR sex identification of embryo is very important for bovine embryo transferring. Many sex identification methods using duplex PCR were established according to Sry gene. But the identification process was affected greatly by more primers interaction. In order to decrease the interference from more primers, researchers explored a simple and rapid PCR Method. The sequences of Amelogenin alleles located at both sex chromosomes were downloaded from GenBank. A pair of sex specific primers was designed to span the 63 bp longer insertion sequence in X chromosome. Bovine samples of blood, fibroblasts and demi-embryos were sexed with these primers. Two-temperature PCR cycling program was used in which the extension step was deleted while the denaturizing and annealing steps were shortened to 1 sec. The results shown ideal identification were obtained and observable amplification were also obtained using even single fibroblast. About 20 bovine embryos were identified by this PCR cycling program and 15 embryos (9 females and 6 males) were transferred. The sexing results were confirmed by the anatomically proven sex after parturition, respectively. The comparison of amplification results between blood samples of bovine and human shows the excellent, specificity to bovine. Thus, a simple, rapid and effective PCR sex identification method was established.
Huabin Zhu, Bo Lin, Jun Chen, Haisheng Hao, Xueming Zhao, Shujing Li, Weihua Du, Tong Qin, Yan Liu and Dong Wang, 2012. Study of a Simple and Rapid PCR Sex Identification of Bovine Embryo. Journal of Animal and Veterinary Advances, 11: 1847-1852.