Equine Embryo Sexing - a Case Study of its Applicability in Romania
Abstract
The interest of sex determination in embryos has an economical and research implication. This method is directly related to the embryo transfer method, which nowadays is considered to be an industry, in both equine and bovine. The purpose of this work was to evaluate the applicability of equine embryo sexing using the polymerase chain reaction (PCR) and to decide if this method is affordable for Romania and yields good results. Five equine embryos have been obtained by natural breeding using transcervical uterine lavage with Ringer Lactate on 6.5 or 7 days. The DNA extraction was performed by using proteinase K and the DNA was quantified by spectrophotometry. Three sets of primers have been used. The double nested PCR was performed, followed by electrophoresis of the amplified samples. From the five embryos obtained, three samples presented a single 445 bp DNA band and no SRY band and they were considered to be females. In two cases, the 445 bp was accompanied by the SRY-1 217 bp and SRY-2 121 bp band, thus being considered to come from male embryos. The embryo biopsy was done in 2 embryos and one biopsied embryo was transferred to a recipient mare. The transfer of the biopsied embryo did not succeed. PCR is suitable for sexing preimplantation equine embryos, but still in order to perform this type of activity at a commercial level, the infrastructure and the development of Equine Reproduction centres it’s mandatory.
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