Summary There has been undertaken a study of mycelia fungus in 42 unique samples of permafrost sediments, collected by the Russian Expedition to the Antarctic within the framework of the project of the International Polar Year “Age of Antarctic permafrost” in 2007-2009 in the ice-free oasises situated along the perimeter of the continent in different geographic zones. With the use of traditional microbiologic methods there was established abundance of fungus, their microfocal zonation over profile and species composition. Parallely, from a part of the same samples (11 samples) after condition selection there was extruded a summarized genomic DNA with the aim of research of uncultivated form of fungal complexes. The consecutive analysis of test samples with the use of molecular-biological methods allowed to obtain data about variety of fungus. Collocation of data obtained by traditional microbiological and molecular-biological methods shows that research of genomic DNA allows to state in subsurface Antarctic samples existence of Ascomycetes and Basidiomycetes, which are difficult or impossible to extrude in standard nutrient solutions. On the other side, in some cases, the method of research the summarized genomic DNA did not allow to show existence in the samples of microspore extremetolerant micromycetes of Penicillium and Cladosporium classes.