Thanks to the bioactive components it contains, royal jelly has many positive effects on health as well as being nutritious. In the study, royal jelly at a rate of 2% (w/v) to raw cow milk whose dry matter was adjusted to 13%, and S thermophilus, Lb. bulgaricus and Bifidobacterium bifidum added probiotic yoghurt (PY_A) with increased functional properties was produced. In the other part, probiotic yoghurt (PY) was produced without adding royal jelly. Physicochemical, microbiological, rheological, sensory tests and color analysis were performed on the samples, and the fatty acid composition and the amount of trans-10-hydroxy-2-decenoic acid (10-HDA) were determined in the yoghurt samples. In the study, the association between the increase in acidity, decrease in carbohydrate level, dry matter level, rheological properties, the viability of probiotics, fatty acid composition, and sensory properties during storage and royal jelly addition was found to be significant (p < 0.05). It was determined that PY_A samples were more acceptable than PY samples in terms of important parameters such as rheological properties, the viability of probiotic cultures, and fatty acid composition. It was determined that the amount of trans-10-hydroxy-2-decenoic acid (10-HDA), one of the most important bioactive components of royal jelly, was above the minimum level specified in the standards in yoghurt samples. Probiotic yoghurts produced with the addition of royal jelly can be a good alternative to innovative functional foods with acceptable product properties.