The Collections of microbial cultures are considered centers of biological resources because they preserve strains of microorganisms that can be used in different microbiological, biotechnological, educational and research studies; therefore is necessary to apply appropriate preservation methods to ensure their viability, availability and protection of their phenotypic characteristics, genotypic and industrial potential. The aim of this study was to establish a collection of microbial strains for use in training and applied in research projects at Coffee for training Center SENA-Regional Caldas. In this study instructors conducted twelve microbial strains from research and trainees using preservation techniques in sterile distilled water and 10 % glycerol those were retained. Preservative was performed in 2 mL cryovials tubes adjusted to an optical density 1 (9x108 UFC/mL) MacFarland scale. The viability and purity was assessed preserved strains, resuming them in selective media for each type of microorganism, in order to observe changes in physiological and morphological characteristics of the cultures and to determine the efficiency of preservation methods evaluated. According to the results obtained in tests, they could be identified fully 12 microbial genera corresponding to Bacillus subtilis, Bacillus pumilus, Candida albicans, Pseudomonas aeruginosa, Saccharomyces cerevisiae, Salmonella sp, Clostridium perfringens, Lactobacillus plantarum, Lactobacillus sp, Pseudomonas putida, Pseudomonas mendocina and Hafnia alvei, likewise positive growth of these microorganisms retain their viability was observed in both purity and preservation techniques used.