Abstract: The cannabis plant is a tall annual crop of economic importance. It is mostly dioecious, but fiber hemp varieties have been bred to be monoecious. Separating male and female plants at early developmental stage is useful due to the influence of gender at agriculturally significant traits. Several experiments have been focused on developing a reliable molecular marker for sex determination in cannabis plants. Our study compares three DNA markers for the detection of male genotype in totally twelve samples of industrial hemp and medicinal cannabis plants. Genotype scoring of SCAR119 marker appeared to be the most reliable, followed by MADC2 and SCAR323, when compared to the observed phenotype of plants. The results confirmed the insights given in previous studies. Research and Polymerase Chain Reaction (PCR) analysis should continue in order to find more advanced DNA markers for sex determination of cannabis plants.