The model plant Oenothera has contributed significantly to the biological sciences and it dominated the early development of plant genetics, cytogenetics, and evolutionary biology. The great advantage of using Oenothera as a model system is a large body of genetic, cytological, morphological, and ecological information collected over more than a century. The Oenothera system offers a well-studied taxonomy, population structure, and ecology. Cytogenetics and formal genetics at the population level are extensively developed, providing an excellent basis to study evolutionary questions. Further, Oenothera is grown as an oil seed crop for the production of essential fatty acids (gamma-linoleic acid) and is considered to be a medicinal plant due to its many pharmaceutically active secondary metabolites, such as ellagitannins. Although Oenothera has been cultivated as a laboratory organism since the end of the 19th century, there is a substantial lack of literature dealing with modern greenhouse techniques for the genus. This review compiles an overview about the growth requirements for the genus Oenothera, with a special focus on its genetically best-studied subsections Oenothera and Munzia. Requirements for greenhouse, field, and agronomic cultures are presented, together with information on substrate types, pest control, as well as vegetative and seed propagation, cross pollination, harvest, and seed storage. Particular aspects like germination, bolting, and flowering induction in taxonomically diverse material are reviewed. Methods recommended are supported by ecological and experimental data. An overview of the possibilities for wide hybridization and polyploidy induction in the genus is given. Germplasm resources are referenced. In summary, a comprehensive guideline for successful laboratory cultivation of Oenothera species is provided.