I want to examine the effect of a drug on a gene expression. But there is a possibility that the increase or decrease in the expression is due to the cell proliferation or cell death.
How can I account for that and study the expression per cell?
Ps, we have PCR, Northern blot, and spectrophotometry equipments.
I feel like I may not totally understand your question, my apologies if that is the case.
However when doing PCR and/or blotting, you should include housekeeper genes whose expression is almost always stable (thus giving a way to normalize data).
So when you look at beta-actin or GADPH expression, these will be constant (you will need to test/look into it if you aren't sure) independent of the given treatment. Then you can use that expression to give an indication of how many cells/DNA/RNA was present.
I am not entirely sure, but a thermofisher video recommends subtracting CT scores to normalize while I would have thought that dividing by the CT scores would be the way to go.
Hope it helps
-Jelle
It depends of the number of cells do you have. If you cultivated cells on small culture bottle, or on a 10 cm2 culture plate, you will have sufficient cells to do a flow cytometry to measure proliferation and death, plus leftover cells for you to extract the total RNA. You will use around 106 cells approximately to do the flow cytometry.
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