Two strategies were utilized to investigate the presence and diversity of psychrophilic and psychrotolerant bacteria in the Iceman (5350-5100 years BP). The first was based on conventional microbiological techniques, i.e. growth of bacterial cells on solid and liquid media. The second was based on the isolation of DNA from small mummy specimens and putative identification of bacteria via DNA sequencing. In addiction, the DNA extracted directly from the Iceman’s tissues was treatedby uracil-N-glycosylase (UNG) to detect repair activity. Microbial cultivation made it possible to identify several bacterial genera such as Ralstonia, Arthrobacter, Methylobacterium, Bradyrhizobium, Bacillus and Pseudomonas. On the other hand, DNA analysis showed that the mummy tissues contained the DNA of Clostridium at an elevated concentration. Thus, there is a sharp distinction between metabolically-active (cultivable) and non-active bacteria. The current research may assist in establishing optimal conservation conditions for the mummy.