What is the minimum number of diseased samples and controls required for RNA sequencing in a rare disease to prove a study? Can we use reference genome as a control?
If you want to detect differentially expressed genes, the minimum is two replicates for each condition. Less than that and you won't be able to compute pvalue associated with the difference of expression. More replicates is better of course. Reference genome is not a control (you'll just map the reads on it) so you'll need healthy samples as controls.
The preferred amount is something like 3 samples of affected and 3 samples of healthy controls if you can possibly swing it, making sure you are sampling from a relevant tissue type of course. However, in rare disease research that isn't always possible. You may only have one affected individual. So the answer becomes "as many as you can reasonably sample" if the numbers are less than 3.
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version 2.3.6
Never use the word "prove" in biology. The actual number of samples required to "prove" something would be "all people that ever have or ever will exist"...but you have neither the budget nor time for that.
please, validate your previous questions: Variant annotations in non-coding regions refer to which exact positions on mRNA? , Align-GVGD scores for SNP values ,