Sex is an important biological variable as many physiological as well as disease processes differ between females and males. The fundamental biological distinction between females and males starts with chromosomal sex, and the establishment of XX and XY cells and tissues. Polymerase Chain Reaction (PCR) is a simple and effective method to easily determine chromosomal or genetic sex of cells and tissues. The goal of this study was to develop a simple multiplex PCR genotyping assay to distinguish XX and XY tissues in sheep. Primers were designed to amplify a fragment of the autosomal gene myogenin (MYOG) and sex determining region on the Y chromosome (SRY). PCR analysis was performed on a variety of genomic DNA samples isolated from fetal sheep skeletal muscle, brain, liver, and placenta, and revealed a single 259 bp band for MYOG in XX females, and a 259 bp band for MYOG and a 167 bp band for SRY in XY males. Amplicons were clearly distinguishable by gel electrophoresis, and their sequences revealed 100% identity to the known ovine MYOG and SRY sequence. The reported multiplex PCR genotyping assay provides a rapid means to distinguish XX and XY sheep tissues using low volume samples.
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Polymerase chain reaction
Sex determining region on the Y chromosome
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The authors like to Asma Omar for her help with the tissue collections and providing the blinded samples. Funding for this work was provided by supported by Agriculture and Food Research Initiative Competitive Grant 2019-67015-29000 from the U.S. Department of Agriculture (USDA) National Institute of Food and Agriculture.
Funding for this work was provided by supported by Agriculture and Food Research Initiative Competitive Grant 2019-67015-29000 from the U.S. Department of Agriculture (USDA) National Institute of Food and Agriculture.
Conflict of interest
The authors declare that they have no competing interests.
All tissues were collected as part of a separate study, and all animal procedures were approved by Institutional Animal Care and Use Committee at Colorado State University, Fort Collins, Colorado, USA.
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Evans, R., Engelhorn, H.J., Winger, Q.A. et al. A multiplex PCR genotyping assay to distinguish XX and XY tissues in sheep. Mol Biol Rep (2020). https://doi.org/10.1007/s11033-020-05701-2