Sensitive sex determination assay applicable to bovine embryos derived from IVM and IVF

A rapid, sensitive method for sex determination which uses nested, allele-specific amplification of bovine zfx and zfy was developed. Accuracy of the sex determination assay was verified with lymphocyte-derived DNA from 24 cattle of known gender. Reliability and sensitivity of the assay in embryonic sex determination were examined by typing duplicate bovine embryo biopsies of 2-8 cells. Sex determination was possible in 38 of 40 biopsies, and agreement between biopsy assays occurred in 17 of 18 cases. Applicability of the sex determination assay was further verified by examining gender of 12 fetuses derived from sexed embryos. Embryonic and fetal gender determinations agreed in 11 of 12 cases with the twelfth yielding an ambiguous result from fetal ultrasound examination.