Technological advances in the last decade have enabled biologists to produce increasing amounts of information for the transcriptome, proteome, interactome and other -omics data sets in many model organisms. A major challenge is integration and biological interpretation of these massive datasets in order to generate testable hypotheses about gene regulatory networks or molecular mechanisms that govern system behaviors. Constructing gene networks requires bioinformatics skills to adequately manage, integrate, analyze and productively use the data to generate biological insights. In this chapter, we provide detailed methods for users without prior knowledge of bioinformatics to construct gene networks and derive hypothesis that can be experimentally verified. Step-by-step instructions for acquiring, integrating, analyzing and visualizing genome-wide data are provided for two widely used open source platforms, R and Cytoscape platforms. The examples provided are based on Arabidopsis data, but the protocols presented should be readily applicable to any organism for which similar data can be obtained.