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We would advise against using the cells from the liquid nitrogen. Cells need to be in their exponential growing phase to give relevant results. Furthermore, this cell line is a stably transfected cell line and requires a selective environment. Cell Growth Medium contains selective agents. The cells need to go through a phase in Cell Growth Medium prior to conducting the assay. This selection is necessary to guarantee that 100% of the cells in the assay are positive for our luciferase-Wnt responsive element construct.