Microbial contamination is a constant problem, which often compromise development of all in vitro techniques. Eight microbial contaminants comprising of four bacteria and four fungi species were isolated from banana (Musa sp.) cv. Grand naine, Ardhapuri and Basrai cultured tissues. The rate of occurrence of bacteria isolates was higher than that of fungi. The bacterial isolate includes Bacillus subtilis, Pseudomonas syringae, Bacillus sp. and Erwinia sp. While fungal contaminants includes Aspergillus niger, Aspergillus flavus, Alterneria tenius and Fusarium sp. This study aimed at investigating possible sources of contamination and means of eliminating these contaminants.