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I'w working with CHO cells. I'm interested in the mRNA level of on gene (the recombinant one). I want to know if a particular condition affects mRNA level.
What I did:
I cultivate CHO cells in shake-flasks for several days.
Each day, I perform RT-PCR with my gene and with b-actine.
For each shake-flask, I calculate the ratio between intensity of my gene and those of b-actine.
Because I perform several experiments with the same culture, I made the replicates in 2 times. Each time, is duplicate. So I have 4 replicates of each conditions.
Due to genetic instability of the cells, I have high differences between replicates, even between the 2 simultaneous replicates.
For example, for one particular day I have
first time condition A: 45% and 37%
second time condition A: 16% and 12%
first time condition B: 35% and 49%
second time condition B: 34% and 19%
My interpretation is that the condition does not affect the mRNA level of the gene.
I want present clearly the results.
Thanks for any suggestions.
