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Cell cycle seems like such a straightforward assay. At its heart, it is a one-color assay and should be a simple protocol to follow. However, as discussed before, fixation and dye concentrations are critical. Once those are optimized, it becomes important to run the cells low and slow in order to get the best quality histograms for analysis — the topic of another blog. Adding the critical CEN and TEN controls will help standardize the assay, and ensure consistency and reproducibility between runs while helping identify non-standard (aneuploid, polyploid) populations from normal ploidy. Trying to isolate and focus on specific components of the cell cycle can be done by addition of specific antibodies or using thymidine analogs. In the end, cell cycle analysis is a simple assay that has a great deal of potential. With work and optimization, a great deal of information about the life of a cell can be extracted.

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