The present work aimed to establish a protocol to minimize the potential effect of in-vitro preservation for 8 months of the two olive cultivars i.e Koronaiki and Kalamata. Three culture conditions (treatments) were chosen to investigate the most suitable one for minimizing somaclonal variations after the process of in vitro preservation. The conditions include MS medium plus mannitol, MS medium plus low temperature and MS medium plus naphthaline acetic acid. RAPD-based finger-printing was employed to determine the effects of the different chemical constituents and other culture conditions on the extent of somaclonal variations after 8 months of in vitro preservation at the DNA level. Four arbitrary primers (OPA-04, -07, -14 and -16) were successfully utilized to achieve such goal. The obtained results showed that Mannitol has more pronounced effect than the other investigated preservation culture and conditions for decreasing DNA alterations.