Abstract
The accuracy of Polymerase chain reaction (PCR) assay in sexing of sheep embryos was assessed in this study. A total of 174 ovine embryos produced in vitro at different stages of development (2, 4-8 cell stages, morula and blastocyst) were sexed. The universal primers (P1-5EZ and P2-3EZ) used in this assay amplified ZFY/ZFX-specific sequences and yielded a 445 bp fragment in both sexes. Restriction enzyme analysis of ZFY/ZFX-amplified fragments with Sac I exhibited polymorphism between sexes, three and two fragments in males and in females, respectively. For verification of accuracy, blood samples of known sex were utilized as positive controls in each test. The mean percentages of sex identification by this method at 2 cell, 4-8 cell, morula and blastocyst were 73.00±5.72, 89.77±3.79, 83.33±8.08 and 79.6±9.09, respectively with the over all male to female ratio of 1:0.87. It is concluded that the ZFY/ZFX based method is highly reliable for the sexing of sheep embryos. (Asian-Aust. J. Anim. Sci. 2003. Vol 16, No. 5 : 650-654)
Highlights
Sheep, with their multifaceted utility, play an important role in the agrarian economy
Embryo sexing by polymerase chain reaction (PCR) and restriction enzyme (RE) analysis have gained importance and offer an accurate and fast method to determine the sex of bovine (Ennis and Gallagher, 1994), porcine (Fajfar-Whetstone et al, 1993) and equine (Peippo et al, 1995) preimplantation embryos
The mean percentage and angle values corresponding to percentages, of embryos were calculated as per the of embryos
Summary
With their multifaceted utility, play an important role in the agrarian economy. Embryo sexing by polymerase chain reaction (PCR) and restriction enzyme (RE) analysis have gained importance and offer an accurate and fast method to determine the sex of bovine (Ennis and Gallagher, 1994), porcine (Fajfar-Whetstone et al, 1993) and equine (Peippo et al, 1995) preimplantation embryos. This method is usually based on the presence or absence of Y-specific amplification (Herr et al, 1990; FajfarWhetstone et al, 1993; Ennis et al, 1999). In this study we assessed the efficiency of PCR and RE analysis in sexing of ovine embryos at different stages of development
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