Okay I got it thanks.
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I have Microarray data with 3 sample each in uninfected_48hrs, infected_48hrs, uninfected_60hrs and infected_60hrs.
#3 sample infected_48h; 3 sample Uninfected_48h; 3 sample infected_60h & sample Uninfected_60h
#First design matrix looks like this:
infected_48h infected_60h Uninfected_48h Uninfected_60h
Uninfected_48h.1 0 0 1 0
Uninfected_48h.2 0 0 1 0
Uninfected_48h.3 0 0 1 0
infected_48h.1 1 0 0 0
infected_48h.2 1 0 0 0
infected_48h.3 1 0 0 0
uninfected_60h.1 0 0 0 1
uninfected_60h.2 0 0 0 1
uninfected_60h.3 0 0 0 1
infected_60h.1 0 1 0 0
infected_60h.2 0 1 0 0
infected_60h.3 0 1 0 0
attr(,"assign")
[1] 1 1 1 1
attr(,"contrasts")
attr(,"contrasts")$sample.levels
[1] "contr.treatment"
# Second design matrix
Uninfected_48h infected_48h infected_60h Uninfected_60h
Uninfected_48h.1 1 0 0 0
Uninfected_48h.2 1 0 0 0
Uninfected_48h.3 1 0 0 0
infected_48h.1 1 1 0 0
infected_48h.2 1 1 0 0
infected_48h.3 1 1 0 0
uninfected_60h.1 1 0 0 1
uninfected_60h.2 1 0 0 1
uninfected_60h.3 1 0 0 1
infected_60h.1 1 0 1 0
infected_60h.2 1 0 1 0
infected_60h.3 1 0 1 0
attr(,"assign")
[1] 0 1 1 1
attr(,"contrasts")
attr(,"contrasts")$SKN_group_int
[1] "contr.treatment"
# Which design matrix is prefer in this scenario first design matrix or second design matrix?
Which design matrix should I used in this case and why? I am new to this design matrix concept. Please do comment and suggest thank you.
in your first matrix, you have all cases separated, then you can compare directly [infected vs uninfected] or [48h vs 60h], or [infected_48h vs uninfected 48h].
In the second case, you are using your uninfected_48h as a base point, so your comparisons depend on that.
So, it depends on what are you asking your data to decide the best design matrix.
Okay I got it thanks.
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Something is not right, here. Please show all prior code, and paste samples of your metadata, if possible. Thank you.
I was curious on which design matrix should I used/based on if I have 12 samples (3 infected_48hr, 3 uninfected, 3 infected_60hr and 3 uninfected_60hr). So I didn't paste the prior code. I think I should used the first design matrix.
Sure, and yes, you can now conduct individual comparisons via
makeContrasts()andcontrasts.fit()