Abstract
Clonogenic assays serve as a useful tool to test whether a given cancer therapy can reduce the clonogenic survival of tumor cells. A colony is defined as a cluster of at least 50 cells that can often only be determined microscopically. A clonogenic assay is the method of choice to determine cell reproductive death after treatment with ionizing radiation, but can also be used to determine the effectiveness of other cytotoxic agents. The following protocol has been modified from a published version (Franken et al., 2006).
Materials and Reagents
Equipment
Procedure
Recipes
Acknowledgments
This protocol has been modified from a published version (Franken et al., 2006).
References
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Dear above friends, could you all see the point of 5th from fixation and staining paragraph, i think it would be wrong one..... is in it....
Although I have never done such experiment,I'd like to share some of my thought with you. From my understanding of this protocol, it should not affect the experiment fundamentally. Should there is any trivial difference, I prefer plate cell first then treat cells since if the other way (treat cells, then plate them) would involve stress the already stressed (treatment) cells one more time (detach the cells). In addition, for the easiness of operation, it is much easier to plate cell first then the other way around. Since for plate cell first case, you just need to aliquot cells from the same master cell suspension, but if treat cell first, then you need to harvest, count and aliquot from different dish of cells. These are the trivial difference I could think of, but shouldn't change the result.Hope it may be helpful.