The Soft Agar Assay for Colony Formation is an anchorage independent growth assay in soft agar, which is
considered the most stringent assay for detecting malignant transformation of cells. For this assay, cells
(pretreated with carcinogens or carcinogen inhibitors) are cultured with appropriate controls in soft agar
medium for 21-28 days. Following this incubation period, formed colonies can either be analyzed
morphologically using cell stain and quantifying the number of colonies formed per well.