The focus of this experiment was to identify unknown bacteria. The identification of unknown bacteria produces benefits for many aspects of the research of microorganisms and helps physicians correctly treat patients. Multiple biochemical tests were performed to provide the fermentation abilities, presence of certain enzymes, and certain biochemical reactions. Qualitative observations were made on the tests, which were compared to unknown bacteria identification key to aid with the identification process. And use of 16S rRNA gene sequences to study bacterial phylogeny and taxonomy has been by far the most common housekeeping genetic marker used for a number of reasons. These reasons include (i) its presence in almost all bacteria, often existing as a multigene family, or operons; (ii) the function of the 16S rRNA gene over time has not changed, suggesting that random sequence changes are a more accurate measure of time (evolution); and (iii) the 16S rRNA gene (1,500 bp) is large enough for informatics purposes. Finally the several amplified parts could be assembled together to have the entire sequence of the complete 16S rRNA. In addition to highly conserved primer binding sites, 16S rRNA gene sequences contain hypervariable regions that can provide species-specific signature sequences useful for bacterial identification. Species identification continues to be a challenge. The development of new methods for this purpose is essential. The acknowledged limitations of the 16S rRNA gene for resolving close interspecific relationships will inspire workers to investigate other genes such as recA, gyrB or GroEL as new targets for molecular assays.