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Colony formation assay

PL Patrick L. Leslie
YC Yvonne L. Chao
YT Yi-Hsuan Tsai
SG Subrata K. Ghosh
AP Alessandro Porrello
AS Amanda E. D. Van Swearingen
EH Emily B. Harrison
BC Brian C. Cooley
JP Joel S. Parker
LC Lisa A. Carey
CP Chad V. Pecot
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After treatment, cells were trypsinized, counted, and 1000 or 5000 cells were plated in triplicate in 6-well plates containing complete RPMI medium and drug, as indicated. Cells were allowed to grow under standard conditions for at least 4 days until colonies were observed. For staining, 1 ml of crystal violet stain (0.05% crystal violet, 1% formalin, 1% methanol in PBS) was added to the cells. Cells were destained in deionized water, and images were taken using an Epson office scanner under film settings. To quantify stain, crystal violet was extracted using 1.5 ml of 1% SDS, followed by absorbance measurement at 612 nm wavelength light on a BioTek luminometer plate reader. Readings were also conducted in triplicate for each sample.

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