The method to analyze both eukaryotic and prokaryotic microorganisms without preliminary microbial information of sample seemed to be useful not only for research and investigation of microorganisms but also for industry using microorganisms. In the present manuscript, preparation of a new DNA primers, new reference database for 18S rDNA for our newly developed method [1]-[3], and analyses of eukaryotic and prokaryotic microorganisms in fermentation products were presented. In komekouji, Aspergillus spp., was enumerated to be 46.5 × 10 6 MPN g −1 , and Penicillium spp., was enumerated to be 1.5 × 10 6 MPN g −1 . In dry yeast, Saccharomyces group, were enumerated to be 8600 × 10 6 MPN g −1 . In komekouji-miso, no eukaryotic microorganism was detected, while the other Bacillus spp., was numerically dominant (21.5 × 10 6 MPN g −1 ) as prokaryotic mi- croorganisms, followed by B. subtilis group (4.65 × 10 6 MPN g −1 ), and the other Firmicutes (3.7 × 10 6 MPN g −1 ). The komekouji-miso included lower number of Actinobacteria (0.15 × 10 6 MPN g −1 ), Burkhokderia sp. (1.5 × 10 6 MPN g −1 ), and the other α,β,γ-proteobacteria (0.12 × 10 6 MPN g −1 ). In sake-kasu, both prokaryote and eukaryote were not detected by the method. Present results indicated that using both universal primers for eukaryotic and prokaryotic microorganisms, each groups of prokaryotic and eukaryotic microorganisms were enumerated without any preliminary information nor setting up standard curve, which were required for real time PCR.