Genetic insights: PCR-based sex verification of Limousin bull (Bos taurus) semen

The determination of spermatozoa sex is a strategic step in the development of artificial insemination technology. The method of separating spermatozoa carrying female (X) or male (Y) sex chromosomes using Bovine Serum Albumin (BSA) has achieved gender accuracy rates of 70% to 80% based on pregnancy rates. However, this verification method
requires a lengthy process to determine the success percentage of sexing. Polymerase Chain Reaction (PCR) testing can be utilized for verification in a relatively short time, requiring a pair of primers to amplify the target DNA region. This study aims to verify sexed spermatozoa using a new primer design for the Limousin cattle X chromosome through PCR testing. The results indicate that sperm separation using the BSA column yielded the PLP1 gene (207 bp). This demonstrates that sperm sexing with the BSA column method can effectively separate sperm carrying X and Y chromosomes, which PCR molecularly verifies.