Very little is known about the higher latitude inland biology of continental Antarctica. In this paper we describe the limnology and biology of the Dufek Massif, using a range of observational, microscopic and molecular methods. Here two dry valleys are home to some of the southernmost biota on Earth. Cyanobacteria were the dominant life forms, being found in lakes and ponds, in hypersaline brines, summer melt water, relict pond beds and in exposed terrestrial habitats. Their species diversity was the lowest yet observed in Antarctic lakes. Green algae, cercozoa and bacteria were present, but diatoms were absent except for a single valve; likely windblown. Mosses were absent and only one lichen specimen was found. The Metazoa included three microbivorous tardigrades (Acutuncus antarcticus, Diphascon sanae and Echiniscus (cf) pseudowendti) and bdelloid rotifer species, but no arthropods or nematodes. These simple faunal and floral communities are missing most of the elements normally present at lower latitudes in the Antarctic which is probably a result of the very harsh environmental conditions in the area.