Abstract：The objective of this study was to examine the possibility of using a non-electrophoretic method to detect the TSPY gene for sexing bovine early embryos, which made bovine embryo sexing under farm condition more feasible. Primers were designed to amplify a portion of the TSPY gene and a common gene as an internal control primer. PCR optimization was carried out using a DNA template from bovine whole blood. Furthermore, six embryo samples were diagnosed by this method and the sexing results were contrasted with those of the Loop-Mediated Isothermal Amplification (LAMP) method. The result showed that TSPY was as reliable a sexing method as LAMP. Forty-three morula and blastocyst embryos collected from superovulated donor dairy cattle were sexed by this method, and twenty-one embryos judged to be female embryos were transferred non-surgically to recipients 6 to 8 days after natural estrus. Out of 21 recipients, 9 were pregnant and all delivered female calves. The results showed that the sex predicted accuracy of this protocol was 100%. In conclusion, TSPY gene was a good male specific marker and indicated that a non-electrophoretic method was feasible and accurate to detect TSPY gene for sexing preimplantation bovine embryos.
