I have values of two treatment groups with similar gene target and all 3 groups have the same beta actin value. Treated 1 = 21.00 cq and Treated 2 = 20.50 cq, while both treatments having the same Control = 19.00 cq. From this rough data, is it reliable to say Treated 2 have higher expression than Treated 1 against Control due to lower cq value by 0.5? Thank you in advance
Hi,
It is not feasible to say that treated2 had a higher expression. The experiment could be done again using technical replicates, which would allow statistical analysis to be done to back up the results.
If your R squared value is 99-100% (technical replicates), your primer efficiency is 95-105% (acceptable efficiency), and your biological replicates don't vary relative to controls, then you MIGHT be able to say yes.
But, I'd need to be convinced by the stats.
If I had to place a bet, my money is on "No, there is no difference in expression". But again, I'd need to be convinced by the stats.
Try to add technical replicates and at least 3 biological replicates to get a statistically significant data. However practically with the difference of 0.5 cT you cannot actually comment too much about the expression of gene. A higher number of biological replicates would help in this case.
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