Cyanobacteria play an important role in the primary productivity of many ecosystems and are dominant in non-marine polar environments. Apart from detecting low levels of cyanobacteria-like pigments in the Southern Ocean, little effort has been spent in trying to elucidate Cyanobacteria in Antarctic sea ice. Here, we report the first use of culture, microscope, microarray and molecular techniques to show that marine Cyanobacteria are rare or absent in sea ice. Our infrequent positive signals were most closely related to freshwater Cyanobacteria from neighbouring terrestrial sources, which illustrates our techniques were sensitive enough to find sea-ice cyanobacteria if they were present. It is still possible that minute quantity of marine cyanobacteria may exist in sea ice and do not contribute significantly to the polar marine ecosystems.