Gluconeogenesis is a continual process that is of great importance in ruminants because almost all dietary carbohydrates are fermented to volatile fatty acids in the rumen. In turn, propionate is the only major volatile fatty acid that contributes to gluconeogenesis. Many different techniques and analytical procedures are involved in studying ruminant gluconeogenesis. Glucose kinetics can be examined by single-injection or continuous-infusion isotope dilution techniques with a variety of glucose labels. Correcting for recycling of label is an important consideration. Absorption of glucose from the gut can be measured by combining arterial-venous differences and flow rates of portal blood or can be estimated by determining the amount of glucose plus alpha-glucose polymers passing into the small intestine. Production of propionate in the ruminoreticulum can be measured by isotope dilution techniques. Quantitating the conversion of propionate to glucose requires the use of [carbon-14]propionate with careful corrections for propionate carbon entering the citric acid cycle before incorporation into glucose; The same fundamental techniques used with propionate are required to quantitate the contributions of amino acids and other precursors to glucose. In vitro studies of gluconeogenic enzymes, and cellular, tissue, or organ preparations provide valuable insights into the gluconeogenic processes and controls but must be validated by in vivo experiments. Progress has been considerable in understanding some aspects of ruminant gluconeogenesis, but many more studies will be required to obtain a complete understanding.