Genetic and epigenetic stability of calli were investigated with RAPD and CRED-RA techniques, respectively. Mature embryos of barley (Hordeum vulgare cv. Zafer-160) were cultured in callus induction MS (Murashige and Skoog) medium supplemented with 1 mg/L 2,4-D and maintained on same medium for 24 weeks. During RAPD analyses 16 primers of tested 20 random primers, produced 103 amplification products. Twenty-five of them were common in both tissues however 24 were, mature embryo and 54 were calli specific. Three primers, which produced best amplification products during RAPD analyses, were used,for CRED-RA analyses and they produced 17 amplification products. There were 8 different amplification products. Culture conditions cause genetic variations and there are also evident cytosine methylation alterations.