I am going to study the expression level of several target genes using qPCR.
In my experiment, I expose microalgae culture to heat stress. then after I harvest the culture in different time points for RNA extraction and ...
the comparison can be done in two different ways.
A) comparing the expression levels of each time point with the expression levels before treatment (zero hours).
B) comparing the expression levels of treatment at each time point with the control at the same time point.
Which one is more accurate?
You have time and treatment as two experimental factors.
Analyze your data (dCt) using a two-factorial model.
Hi,
If you have a endogenous gene as control, you can to use a model for relative expression of your genes using ddCt and you should be to compare each point of treatment respect to zero time (before the stress).
Dear Lesther Lopez-Cruz,
How about comparing each point of treatment respect to corresponding control?
This means to include the microculture-ID as another orthogonal factor in the model. It depends on the variance structure is this is advantageous. If you have many different time-points, a mixed model with ID as random factor might be better. I would discuss this with a local statistician.
Thanks for your guide Prof. Wilhelm,
In the experiment, the culture will be grown at high temperature and control temperature and then in 7-time point will be harvested for gene expression analysis.
since the cultures only affected by time and treatment I assumed that the repeated measure ANOVA can be used to compare the significance of difference.
- Badges
- Science method