Hi All!
I have been given a number of RNA samples for an upcoming RNA-seq experiment, but I've recently discovered that the number of cultured cells used for RNA extraction was not consistent (i.e., each extracted sample consisted of a different number of cells). I'm worried that using RNA extracted from different cell densities will have an effect on downstream differential expression analyses. I've searched around a decent amount, but am having trouble finding any solid information regarding my situation. Before I move forward with this experiment, I'm wondering:
I have been given a number of RNA samples for an upcoming RNA-seq experiment, but I've recently discovered that the number of cultured cells used for RNA extraction was not consistent (i.e., each extracted sample consisted of a different number of cells). I'm worried that using RNA extracted from different cell densities will have an effect on downstream differential expression analyses. I've searched around a decent amount, but am having trouble finding any solid information regarding my situation. Before I move forward with this experiment, I'm wondering:
- How concerned about this should I be?
- If I move forward with sequencing, should I try to control for cell number by adding it as a covariate in my analyses (if I can get this data)?
- Are there any other potential solutions that I should consider?
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