Single Cell PCR Using Y Chromosome Specific Repeat for Sexing

Abstract

Aims: Embryo sexing is one of the important ways for sex selection of offspring. This is a potential method to considerably improve animal breeding and the efficiency of dairy and meat production. A novel repeated sequence specific to male cattle has been identified and named S4. S4 is a 1.5 Kb repeating unit contains various internal repeated sequence. S4 is localized on long arm of the Y chromosome in the region near to ZFY genes. The objective of this study was to establish a simple, sensitive, reliable, reproducible and cost effective PCR based technique for sexing. Study Design: Case control study. Place and Duration of Study: Sample: whole blood samples of male and female cattle. Medical Biotechnology Dept. National Institute for Genetic Engineering and Biotechnology, Tehran, 14965/161, Iran, between June 2010 and July 2011. Methodology: Genomic DNA was extracted from the whole blood samples of male and female cattle. PCR and single cell PCR were performed using specific primers for this region. Results: By this PCR based methods we could differentiate between female and male genomic DNA. Conclusion: With this technique we can distinct male from female using as little as 0.1pg DNA. Using this method we could determine the sex of embryo (4 blastomers). This method may optimize for the quantitative detection of Y chromosomes in semen.