The bamboo is a plant that possesses multiple uses with an enormous economic, social, cultural and environmental impact. The present research work was developed with the objective of determining the effect of different disinfection methods in the establishment and growth in vitro of Bamboo (Guadua angustifolia Kunth.). Two disinfection methods were applied: a simple method based on the use of sodium hypochlorite at 1.0%, 2.0% and 3.0% during 20 minutes, and a double method in which sodium hypochlorite was employed at 2.0% during 5, 10 and 15 minutes with repetition from the disinfection at 24 hours. At one, two and three weeks the following variables were evaluated: percentage of contamination, phenolization , death and budding , number and length of the buds for explant. A design totally randomized with one way analysis of variance and means comparison test of minimum significance difference were applied. The results demonstrated that the method of double disinfection based on the use of sodium hypochlorite at 2% during 5 minutes with repetition at 24 hours is the most appropriate for the in vitro establishment of primary explants of Guadua angustifolia Kunth. Key words: bamboo, plant tissue culture, sodium hypochlorite, contamination, primary explants