Rapid sex determination of preimplantation bovine embryo using direct-PCR from a single blastomere

Introduction: Sex control plays an indispensable role in livestock farm management. It enables farmers to quickly respond to the needs of herd rearrangements such as size and composition, and stably maximize the economic value, particularly on long gestation period animals as bovine. Despite being an invasive method, genetic examination of blastomeres isolated from the preimplantation embryos provides a superior accuracy method for bovine sexing, particularly with a highly sensitive molecular technique such as PCR. Currently, this technique is commercially expanding due to many improvements in embryo technology. In this study, we attempted to reduce the embryonic invasiveness and develop a simple, fast, and reliable sexing procedure using direct amplification of Y chromosome-specific marker from only one blastomere. Methods: The bovine sexing was carried out using direct PCR of Y chromosome amplicon from single blastomeres biopsied from sex-known somatic nuclear transfer technology (SCNT) embryos (male) and enucleated metaphase II nucleus (female). The intensity of the amplified products was also optimized using the PCR additive. Results: Utilizing the protocol resulted in the success of sex typing for all tested samples in this study that are comparable with genomic DNA amplification. Conclusion: Our result could serve as a fast procedure for bovine embryo sexing and would be an effective tool for sex ratio control in the husbandry industry, animal embryo research, and endangered animal preservation.