DE analysis with limma partial replicates
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ea1402 ▴ 20
@ea1402-8294
Last seen 8.1 years ago
United States

I would like to analyze RNAseq with 6 conditions using limma/voom. I have 6 biological replicates for control, however for the other 5 conditions (say C1,C2,C3,C4,C5) I have no replicates. Ideally, I would like to do a variance estimation using the 6 replicates for the control, and using it compare any two condition C1 vs C2. Does the standard workflow takes care of this?

 

thanks in advance

limma voom • 920 views
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Aaron Lun ★ 28k
@alun
Last seen 4 hours ago
The city by the bay

Yes, provided you specify the design matrix properly, i.e., in a one-way layout where all replicates for the control condition are in the same group. However, even though limma will handle it, I feel obliged to point out that your set-up is far from ideal; there's no way to spot aberrant behaviour within each of the other conditions if you've only got one replicate for that condition. Thus, you should probably take your DE results with a grain of salt, especially when you're comparing between two conditions.

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