The prohibition of Cannabis sativa has stifled modern research, resulting in underdeveloped biotechnological tools and difficulties replicating existing protocols. With the growing market, in vitro practices are crucial for maintaining Cannabis germplasm and developing biotechnologies. This thesis re-evaluates the basal salts and plant growth regulators (PGRs) commonly used in existing Cannabis protocols and tests different growing orientations to improve multiplication rates and health of drug-type cultivars. The results indicate that Driver and Kuniyuki Walnut (DKW) basal salts are superior to Murashige and Skoog (MS) basal salts for multiple drug-type cultivars, improving growth and multiplication rates by 1.96x and 1.5x, respectively. Screening four common PGRs showed no significant improvement over plain DKW basal medium, however, early rooting increased plant growth. Finally, different orientations did not improve growth compared to the control. This thesis provides a foundation for Cannabis micropropagation and emphasizes the need for broader ranges of cultivars when optimizing protocols.