Aims: The aim of this study was to isolate and identify microorganisms in crude oil polluted soils in Eastern Tanzania. Study Design: Isolation and identification of microorganisms using conventional and molecular techniques. Place and Duration of Study: Department of Molecular Biology & Biotechnology, University of Dar es Salaam; October 2012 to June 2013. Methodology: Soils were collected from Mlalakua (ML), University of Dar es Salaam (UDSM) and Ubungo (Ub) local garages in Dar es Salaam, Tanzania. The selective isolation of microorganisms was done based on colony morphological appearance on agar plates and Gram reaction. Three isolates (one from each site) were subjected to molecular methods for the identification purposes. Phylogenetic analyses of bacterial 16S rRNA gene and fungal ITS region sequences were employed in this study to identify and determine the evolutionary relationships of the ML, Ub and UDSM isolates. Results: The 16S rRNA analysis revealed that the ML isolate belonged to the genus Klebsiella, while the ITS analysis on Ub and UDSM isolates assigned them to the genus Trichosporon and Candida, respectively. Conclusions: From these findings, further studies are recommended to assay for the biodegradative potentiality of the identified isolates for bioremediation of crude oil contaminated soils.