| The amelogenin(AMEL) gene in goats is located on the homologous region of both X and Y chromosomes. Based on AMEL-X and AMEL-Y genes with varying degrees deletions, the nested primers were designed. Several aspects were studied including the accuracy of nested PCR primers, the method of amplifying ultratrace genomic DNA, nested PCR amplification of embryonic DNA sample and the transfer of embryos. The purpose of this research is to construct sex identification system on early embryos of goats by nested PCR. The results are as follows:1. The blood genomic DNA of goats was amplified using AMEL gene external primers and inner primers, respectively. Then PCR products were separated by electrophoresis in 1.5% agarose gels and visualized by ethidium bromide staining; external primers were utilized to amplify a 349 bp fragment from X chromosome in female goats, and a 349 bp fragment from X chromosome and a 289 bp fragment from the Y chromosome in male goat, inner primers were utilized to amplify a 311 bp fragment from X chromosome in female goat, and a 311 bp fragment from X chromosome and a 251 bp fragment from the Y chromosome in male goat, respectively. Forty-six blood DNA samples were detected and the result the accuracy in accordance with the actual state was 100%( 22F/22, male:24/24).2. Ten pg genomic DNA (approximately 3 cells) of 6 goats(3F, 3M) were amplified and a fragment from X chromosome in female goat, and a fragment from X chromosome and a fragment from the Y chromosome in male goat.3. Eighty-six usable embryos were obtained by superovulation; thirty-six embryos of goats were detected and 33 embryos were amplified successfully, The sex ratio of the embryo was 16F/17M. 50 embryos were tansfered into recipient goats and 27 lambs were born, The pregnancy rate was 54% (27/50). The sex ratio of offsprings was 12F/15M. There was no significant differences (P>0.05) of sex ratio between sex identification and embryo transfer.
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