Hello,
I am quantifying bacteria exposed or not to a certain chemical at specific time points.
How can I compute the results in a relative manner? That is: what is the formula for a ∆∆Ct analysis of bacteria?
Relative quantification is normally given for a gene of interest over one or more reference genes by calculating the cycle difference between the exposed and not-exposed genes and then between GOI and REF.
For bacteria, would these be acceptable:
∆Ct = Ctt – Ct0
∆∆Ct = ∆Ct (exposed) – ∆Ct (control)
where t is a specific time (0,1,2,4,24 h post-exposure) and 0 is t=0. The fold increase would then be 2–∆∆Ct.
Is this correct?
Thank you
In principle yes, but where is the time in your model? As I understood, you should model the change in dCt over time, and test for a difference in a meaningful model parameter.
The point is to check the growth of bacteria at t= 0,1,2,4,24 h and compare the growth of the exposed to that of the control.
What has growth to do with the dCt values? If you want to analyze the growth, use growth models.
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