Xueming  Zhao, Bo  Lin, Linbo  Zhang, Haisheng  Hao, Weihua  Du, Huabin  Zhu, Jiaming  Guo, Dong  Wang, 
    A Study of a Method to Assess the Purity of Sorted Bovine Semen Using Rapid Single-Sperm Sexing PCR,
    Journal of Animal and Veterinary Advances,
    Volume 10,Issue 6,
    2011,
    Pages 750-756,
    ISSN 1680-5593,
    javaa.2011.750.756,
    (https://makhillpublications.co/view-article.php?doi=javaa.2011.750.756)
    Abstract: Sort reanalysis using flow cytometry is the most common method for determining the purity of X or Y enriched semen. The high cost of this technique (including the required expensive, proprietary machine) limits efforts to improve the technique and to promote develop applications for the sorted semen. In this study, the sperm sex (the presence of the X or Y chromosome) was identified by both rapid PCR and flow cytometry reanalysis. The rapid PCR results showed that the percentages of X and Y sperm were 48 and 52% in unsorted semen, 92 and 8% in X-enriched semen and 17 and 83% in Y-enriched semen, respectively. Reanalysis of the DNA content of the sorted samples revealed that the X and Y sperm frequencies were 92 and 8% in X-enriched semen and 15 and 85% in Y-enriched semen, respectively. The sex ratio of unsorted semen analyzed by PCR did not significantly deviate from the expected ratio of 1:1 and there was no significant difference between the sex ratios of sorted semen samples determined by PCR and flow cytometry reanalysis. These results indicate that we have established an effective, reliable and rapid PCR method to verify the purity of sorted semen. This method should contribute greatly to the improvement of sperm sorting techniques and the development of applications for sorted semen.
    Keywords: Single sperm;sex typing;rapid PCR;bovine semen;cytometry;sorted semen