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Sex Determination in Ovine Embryos Using Amelogenin (AMEL) Gene by High Resolution Melting Curve Analysis

Author(s): Yao Jing Yue | Jian Bin Liu | Ting Ting Guo | Rui Lin Feng | Jian Guo | Xiao Ping Sun | Chun E. Niu | B.H. Yang

Journal: Journal of Animal and Veterinary Advances
ISSN 1680-5593

Volume: 11;
Issue: 16;
Start page: 2948;
Date: 2012;
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Keywords: B.H. Yang | amelogenin gene | sexing | high-resolution melting | blood | China

In the study, researchers have established and tested the reliability of a method for sex determination of ovine embryos using the AMEL gene by melting curve analysis of PCR amplification. It can be carried out in a regular laboratory or under farm conditions within 1.5 h for 96 samples. The PCR amplicons of 99/99 and 99/54 base pairs produced from female and male sheep, respectively are easily distinguished by both melting curve analysis and gel electrophoresis. The specificity of the method was earlier demonstrated by testing 9 blood samples from small-tailed sheep (5 males and 4 females). No amplification failures and very high agreement between genotypic and phenotypic sex was found (9/9). The sensitivity of the AMEL sexing assay was established for values >10 pg ovine genomic DNA. Forty five biopsied embryos were transferred into 22 recipient sheep on the same day that the embryos were collected and sex of the kid was confirmed after parturition. About 17 kids of predicted sex were born. The sex, as determined by PCR corresponded to the anatomical sex in all cases. To the knowledge, this was the first time that sex determination using the amelogenin gene was performed in ovine embryos by melting curve analysis.
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