The main factors that influence formal sample size calculations for RNA-sequencing experiments are: depth of sequencing, coefficient of variation, magnitude of differential expression detected, false positive rate and power. Assuming average read depth of 20, equal within group coefficient of variation of 0.4, an effect size of 2 fold, 0.05 false positive rate and 80% power, the number of subjects per group required is 7.1 The investigators therefore aim for a minimum target of ten samples per group.