The goal of the experiment is to analyze cell cycle after adding a drug.nthanks
In order to provide statistical analysis, you should run in triplicate, the negative and positive control, and the cells treated. If you are going to use a kit for cell cycle analysis, following the manufacturer's instructions you should have no problems. If you are going to stain only with PI, usually 1.105 cells are accepted.
In order to provide statistical analysis, you should run in triplicate, the negative and positive control, and the cells treated. If you are going to use a kit for cell cycle analysis, following the manufacturer's instructions you should have no problems. If you are going to stain only with PI, usually 1.105 cells are accepted.
I agree with Ana and would like to add that you need to collect enough events to accurately measure each sub-population. Try at least 10e4 events. Look at the S and G2 populations to make sure they are well represented.
i am using six well plate so i am using duplicate not replicate. would this work? two wells for treated and two positive control and two wells negative control.
You need to consider the eventual statistical analysis of your data. Duplicates gives you more cells to stain (and should be plenty for a small set of stainings), but duplicates are no good for any statistical analyses if you analyze each well separately. n = 3 is really a bare minimum. Better, off course, is repeated experiments, which means duplicates are fine if results are going to be averaged and later combined with the results of replicate experiments.
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