I have been doing some digging to understand how to use my log amplified histogram from Flow Cytometry output and calculate a 1C value for my plant in question. I have clear peaks but means that do not correlate to each other as they should, i.e. a peak which would be the 4C of the other does not have double the mean value.
I have attached an image of my results. There are two tables in it, one in the graph with the peaks and one below it.
1) Could you tell me if and how I can use the log3 mean values of my histogram to determine the C value?
2) What does the Mean-x column of the table below the histogram mean? There are values that show me a relationship of double for the two peaks that are 2C and 4C. (Peaks 2 and 4 which would be 2C and 4C respectively).
The reference peak is the tallest one and the genome size in question is the next one to its right, Peaks 2 and 3 as labelled on the histogram.
Kindest regards,
Charlotte Moore
This is just some general answers on flow data... (I have no experience in using flow for DNA analysis, so I have no idea how you are suppose to do the calculations.)
1. Do you want mean or median? The median value will reflect the "peak", the mean may or may not depending on the shape of the peak. (If you look at the PK4, mean-x value, it is double that of PK2.)
2. The mean-x value is the mean of the FL1within the gate.
3. You gate for P4 is over two peaks, might want to redo that one.
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