Imagine the following - having lines of Arabidopsis overexpressing certain protein I want to measure the activity.

From one line I will collect leaves from several plants separately (biological replicates). These I will grind, and split by certain amount (primary technical replicates). After extraction I will measure the activity in triplicates (secondary technical replicates).

The question is, if I had say 72 numbers (8 biological x 3 1° technical x 3 2°technical), what do I do with them? Should I just make average from all of them? Or should I first make average from the activity measurement, from those averages make average of the 1° technical replicates and from those the average of biological replicates?

Those two should yield the same result if the number of replicates will be the same for all lines, right?

Or is there other way to treat the samples?

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