Nitrogen fixation by symbiotic and non-symbiotic bacteria can be a significant source of nitrogen in cropping systems. However, contributions from non-symbiotic nitrogen fixation (NSNF) are dependent on available carbon in the soil and environmental conditions (soil moisture and temperature). In Australia, measurements of NSNF have been made in the field by quantifying nitrogenase activity. These studies have included determinations of the moisture and temperature requirements for NSNF and for crop residue decomposition that supplies carbon to NSNF bacteria. Other studies have determined the N input by NSNF using N budget calculations. These data together with information about carbon supply and environmental conditions were used to estimate potential NSNF in the cropping zones of southern Australia. Using the ArcviewGIS Spatial Analyst (v3.1), maps of Australia showing estimates of NSNF in different cropping zones as determined by rainfall and temperature or carbon availability were generated. In Western Australia (represented by Wongan Hills) and South Australia (represented by Avon), where summers are dry, estimates of NSNF were generally low (10–15 kg N/ha from January to June) due to limitations of soil moisture. In New South Wales, particularly in the north where summer rainfall patterns develop (represented by Gunnedah), the warm, moist conditions produced higher estimates of NSNF (totaling 32–38 kg N/ha from January to June). In this region, the majority of estimated NSNF occurred in January and February leading to the depletion of carbon supplies and reduced NSNF in autumn (March–June). Information about potential supplies of N from NSNF across the cropping zones should be useful for researchers to select and study areas that are most likely to benefit from NSNF. It should also help agronomists and extension officers explain changes in N status within paddocks or within specific farming systems and to provide more accurate advice on N fertiliser requirements, particularly in low-input farming systems.