Hi, I have 5 samples, namely the following: (1) Not transfected, untreated (2) Transfected but untreated (3) Transfected, treated, analyzed after 5 mins (4) Transfected, treated, analyzed after 60 mins (5) Transfected, treated, analyzed after 4 hrs [By transfected I mean a particular vector is present, and treated means treated with an antibody]

I do not have any replicates for any of the conditions. **I am looking to perform an Anova test using edgeR**, to see the gene expression at different time points wrt sample 2 (as given).

**Could anyone tell me the edgeR code I should run to do the test?**
Normally, I define the groups, normalize the library, create a design matrix with the group and normalized DGEList, followed by estimating dispersion and performing the GLMQL tests.
I am confused as to how to proceed in this case,as I have no replicates. I don't know how to define the groups or estimate dispersion in this case.

**I would be very grateful if someone could help me with the edgeR codes.
Thank you**

Hi Lun, I want to ask a question. Why do you say that the model above cannot compare specific time points? Is it because when you use

`coef=3`

to compare it with reference, both the time and transfect condition are different, so that it's impossible to say the DE is from time or transfection? Thank you