Hi all, I have one question regarding zebrafish embryos. After a specific treatment, I collected embryos 3 days post-fertilization. Due to the high mortality rate, I could collect 3 cohorts as being 10 embryos in each cohort. I will extract RNA and then conduct qPCR. In the related articles, I checked, saw that cohorts generally consist of 20-30 embryos but I have 10 for each cohort. Do you think does it affect gene expression patterns, especially significance values? I know that by extracting RNA from 10 embryos, I decrease the biological variability. However, I am not sure about its further outcomes. By the way, I have also 10 embryos in my untreated/control cohorts and convert the same amount of RNA into cDNA, surely. Any thoughts/comments are appreciated.