The aim of the present study was to establish the protocol for micropropagation of blackberry cultivar “Reuben”. In aseptic conditions using stereomicroscope, shoot tips were isolated from axillary buds and placed on medium (EM) to establish aseptic culture. The effect of different sugars on PM2 medium in the two variants of 30 g / L sucrose and 20 g / L glucose + 10 g / L sucrose was tested. In order to establish optimal variant of growth regulators 4 variants were experimented, 6- benzylaminopurine (BAP) in concentration of 0.3 mg/L and 1 mg/L and meta-Topolin (mT) in same concentrations. In vitro rooting was carried out in two experimental variants, on IAA rooting medium and on hormone free rooting medium. The greatest number of shoots with average of 2.6 were produced in medium containing 1 mg/L BAP and the maximum shoot length of 8.9 was produced in medium containing 0.3 mg/L BAP. The rooting percentage was higher (61%) on hormone free rooting medium but the acclimatization percentages for both variants were very similar (92% and 95%).