I wanted to know how to decide the treatment time for clonogenic assay with chemo drugs. Would you choose the time when cell death is first noticed with the respective drug treatment or is there a standard treatment time that is used for the assay in general?
Treatment time with the chemotherapeutic drug of interest may depend on various factors, such as the purpose of a clonogenic assay (e.g., to see cytotoxicity to a single agent or in combination with other agents), available quantity and concentrations of drugs, plating efficiency of cell lines, and size of dishes or flasks used for the assay. To obtain survival curve, several dose points may be tested at which colonies after highest dose need to be countable. It will be better if a negative control (a clonogenic assay for a solvent, e.g., PBS or DMSO) can be included as a parallel experiment.
Ruby,
At first, calculate IC50 of that particular drug for your cell line. Then you can treat different concentration of that drug for 8-14days.
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