Bacteria with the ability to grow on nitrogen-free media and fixing atmospheric nitrogen were isolated from different locations in Egypt. Isolates were identified as Azotobacter chroococcum according to their morphological and physiological properties. Isolates were ovoid to rod shaped occurs in pairs, form cysts. Analysis  with Random Amplified polymorphic DNA-Polymerase Chain Reaction (RAPD-PCR) to compare the similarity pattern of the selected Azotobacter isolates and reference strain was used as an identification tool.Physiological characteristics of A. chroococcum such as acetylene reduction activity (nitrogenase enzyme) and production of some plant growth promoting substances such as Indol acetic acid (IAA), Hydrogen cyanide (HCN) and Sidrophore production were also studied.