This study was carried out during the autumn season of 2021-2022 to establish a detailed protocol for the micropropagation of Jacaranda mimosifolia trees. The best results in terms of eliminating contamination (0.0%) were obtained by soaking seeds in Clorox (NaOCl) solution for 9 or 12 min. The highest seeds survival rate (100%) was obtained when sterile seeds were planted in ¾ strength MS medium + 30 g/l sucrose + 6% agar, and pH 5.7. The results showed that ½ strength MS was a suitable medium for the next multiplication stage, because it later gave the highest number of shoots, and chlorophyll A content. On the other hand, the ¼ strength MS medium was suitable for the rooting stage because it resulted in the best root length and shoot length. In terms of shoot length and fresh weight were obtained with 2.0 mg/l 6-BAP + 0.0 mg/l NAA, while 4.0 mg/l 6-BAP + 0.0 mg/l NAA gave the highest number of shoots, and 1.0 mg/l 6-BAP + 0.0 mg/l NAA resulted in the best flavonoids content. The highest root number and dry weight were obtained with 2.0 g/l AC + 0.5 mg/l NAA, while the longest shoots were obtained with 0.0 g/l AC + 0.1 mg/l NAA. In the acclimatization stage, the best plant height was obtained with Peat moss + Perlite at a ratio of 2:1 (v/v). The survival percentage (100%) was equal in both of the tested acclimatization media, viz., Peat moss + perlite at 2:1 (v/v) and Peat moss + perlite + vermiculite at 2:1:1 (v/v/v). The economic feasibility and production costs were calculated after determining the appropriate media for the different micropropagation stages of J. mimosifolia.