I am planning to sort transgenic hESC lines based on reporter lines fluorescent markers, to see if FACs sorting of progenitors results in greater Da yields. For my control study I have included a scrambled gRNA subculture. I wondered if I would have to make all subcultures, even those which will not be sorted (for comparison of sorted vs unsorted) reporter lines as a control?

Alternatively, if this is not necessary i will just compared transgenic sorted culture outcomes with original hESC?