Fetal sex determination in Holstein cow using non-invasive technique Cell-free fetal DNA (cffDNA) in plasma derived from pregnant cows

Discovery of accurate, easy and economic embryo sex determination in the dairy cattle industry assumed highly particular importance. CffDNA is novel potential material to determine the sex of the fetus in early gestation period has not imposed negative effect on embryo health. In this regard, in overall, 16 Holstein cows experiencing eight weeks of gestation and 4 non-pregnant Holstein cows, two bulls (negative control) were chosen as candidates for this present experiment and subsequently that plasma was separated to extract ccfDNA. Qiagene commercial kit was used to extract. DEAD box protein gene-based primers for this study based on both chromosomes X and Y and past articles were selected and were synthesized reported. Polymerase reaction and the required conditions were optimized using proposed standard. The results of electrophoresis PCR patterns male fetus’s two bands and one band of female fetuses showed. Monitoring of birth of each gender embryos under the test after the birth of calves showed compelet match the molecular results re-confirmed