The accurate determination of the spatial trends on the variability of the gene pool of a species is essential to elucidate the underlying demographic-evolutionary events and, ultimately, to unravel the microevolutionary history of the species or population under examination. In this work, we present a new software tool called GenoCline, which is essentially devised to assist in detecting genetic clines from allele and phenotypic frequency data, and even from genome-wide data. This program package allows identifying the geographic orientation of clinal genetic variation through a system of iterative rotation of a virtual coordinate axis. At the same time, GenoCline can also be used to carry out complementary statistical analyses aimed at clarifying the potential origin of the genetic clines found, among which stand out spatial autocorrelation, isolation by distance, centroid method, multidimensional scaling and Sammon projection. Among the main advantages of this software are those related to ease in data entry and potential interconnection with other programs. Data entry is user-friendly. Genetic frequencies and geographic coordinates can be easily entered in spreadsheet table formatting, while genome-wide data can be imported in Eigensoft format. Genetic frequencies can also be exported in a format compatible with other programs dealing with population genetics and evolutionary biology. All illustrations of results are saved in eps format so that there will be high quality and easily editable vectorial graphs available for the researcher. GenoCline is implemented in Java, so it can be used with different operating systems.