PCR-based sexing of caprine embryos using Sry/Aml-X duplex PCR and Sry/DVEPC041 duplex PCR

Abstract

This study presents the development of a PCR-based method for sex determination of caprine embryos by Sry and Aml-X gene detection. DNA sequencing of the PCR products amplified from the target loci confirmed that these amplicons were indeed amplified from portions of the caprine Sry and Aml-X genes. Furthermore, the primer sets were found to be specific for sexing of caprine of various breeds. An alternative PCR-sexing protocol was also developed to verify embryo sexing results and it was a pit-stop using Sry and DVEPC041 primer sets. For the pitstop PCR protocol, the Sry primer was best added after the first 6 cycles to obtain bands of equal intensity at 116 bp and 160 bp when identifying the male sex. However, application of the same pit-stop PCR protocol using genomic DNA of various caprine breeds also produced acceptable and accurate sex identification despite showing slight diversion of the expected result.